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Actinobacillus pleuropneumoniae

Table of Contents
  1. General Information
    1. NCBI Taxonomy ID
    2. Disease
    3. Introduction
    4. Microbial Pathogenesis
    5. Host Ranges and Animal Models
    6. Host Protective Immunity
  2. Vaccine Related Pathogen Genes
    1. sd
    2. APP7_1016 (Protective antigen)
    3. ApxIA (Protective antigen)
    4. apxIA (Protective antigen)
    5. ApxIIA (Protective antigen)
    6. apxIIA (Protective antigen)
    7. CP (Protective antigen)
    8. Lipopolysaccharide (Protective antigen)
    9. apxIA (Virmugen)
    10. ApxIC (Virmugen)
    11. apxIIA (Virmugen)
    12. ApxIIC (Virmugen)
    13. apxIIIB (Virmugen)
    14. apxIIID (Virmugen)
    15. ApxIVA (Virmugen)
    16. ureC (Virmugen)
  3. Vaccine Related Host Genes
    1. A2M
    2. Ighg1
    3. Ighv1-9
  4. Vaccine Information
    1. A. pleuropneumoniae ApxIa and ApxIIa protein vaccine
    2. A. pleuropneumoniae ApxIA protein vaccine
    3. A. pleuropneumoniae DNA vaccine pcDNA-apxIA/pcDNA-apxIIA
    4. A. pleuropneumoniae HS93C-Ampr
    5. Actinobacillus pleuropneumoniae apxIA mutant vaccine
    6. Actinobacillus pleuropneumoniae ApxIC mutant vaccine
    7. Actinobacillus pleuropneumoniae apxIIC and apxIVA double deletion mutant vaccine
    8. Actinobacillus pleuropneumoniae apxIIIB and apxIIID double mutant vaccine
    9. Actinobacillus pleuropneumoniae HS93Tox-/pIG-T1K vectored vaccine
    10. Actinobacillus pleuropneumoniae ureC and apxIIA double mutant deletion vaccine
    11. HS93Tox-/pIG-T1K
    12. PleuroStar APP
    13. Porcilis APP
    14. rS.C-APP-ApxI/ApxII
    15. rS.C-APP-ApxIIA
  5. References
I. General Information
1. NCBI Taxonomy ID:
715
2. Disease:
porcine pleuropneumonia
3. Introduction
The bacterium Actinobacillus pleuropneumoniae (App) was previously called Haemophilus pleuropneumoniae and there are at least twelve different strains, some of which produce no disease and are non pathogenic, but others cause very severe disease. Strains 1, 5, 9, 11 and 12 are highly virulent and strains 3 and 6 are very mild. The organism is carried in the tonsils and respiratory tract and the incubation period is very short, from as little as 12 hours through to three days. It is transmitted by droplet infection between one pig and another. The organism may survive in discharges, serum etc. for up to 5 days. Contact with dead stock is therefore important from biosecurity. It dies quickly if dried, but it may persist in water for 20 days or more. App can survive in the lungs and tonsils for long periods of at least 4 months. It is probably airborne for only 5 to 10 metres. Disease is dose dependent i.e. the more bacteria the pig is exposed to the more severe will be the disease. Infection is spread from one pig to another by nose to nose contact. Pigs may be infected with different serotypes simultaneously. PRRS and EP can make the disease worse. In a naïve herd up to 30% of animals may be affected. When App attacks the lungs the toxins produced cause severe damage to the tissues which turn blue to black (necrosis) with extensive pleurisy. The chest cavity rapidly fills up with fluid.

The organism may affect the pig from weaning through to slaughter but usually the age is from 8 to 16 weeks, once maternal antibody has disappeared. Sudden death is often the only sign with blood and froth discharged from the nose. In the live pig a short cough may be heard with signs of severe breathing difficulties and blueing of the ears. Badly affected pigs are severely depressed. Body temperature is often high. Death is due to a combination of heart failure and the toxins produced by the organisms (ThePigSite Pig Health).
4. Microbial Pathogenesis
Actinobacillus pleuropneumoniae interacts closely with epithelial cells in the lower respiratory tract and toxins produced during infection are delivered directly to the cell surface. This close association between A. pleuropneumoniae and respiratory epithelial cells may impair the binding of specific antibodies to the Apx toxins, resulting in development of necrosis and hemorrhage (Haesebrouck et al., 2004).
5. Host Ranges and Animal Models
Pigs are the main host, and mice can serve as a model of infection (Shin et al., 2007).
6. Host Protective Immunity
Protection is mediated by antibodies and mucosal immunity (Shin et al., 2007).
1. APP7_1016
  • Gene Name : APP7_1016
  • Sequence Strain (Species/Organism) : Actinobacillus pleuropneumoniae serovar 7 str. AP76
  • VO ID : VO_0011022
  • NCBI Gene ID : 6398034
  • NCBI Protein GI : 190150285
  • Locus Tag : APP7_1016
  • Genbank Accession : CP001091
  • Protein Accession : YP_001968810
  • Taxonomy ID : 537457
  • Gene Starting Position : 1141773
  • Gene Ending Position : 1142414
  • Gene Strand (Orientation) : -
  • Protein Name : putative hemolysin activator protein
  • Protein pI : 7.21
  • Protein Weight : 23153.77
  • Protein Length : 213
  • DNA Sequence : Show Sequence
    >gi|190149269:1141773-1142414 Actinobacillus pleuropneumoniae serovar 7 str. AP76, complete genome
    CTCACTTTAAATTCTCAAGTGACTGTTCAATATTGCGCACATTTAAAATATCGCCTTTTTCAAAGGTAAA
    AGCGGTCAGAGCGTGTAAAGGGGTAAAACGAGGTACTCGACCGTTATCCATCACGACCGTATTACCTATT
    TTTCCGACAATCACGCTTAGCACTAAATTACCGGAACCTAAATCCTGCTCTTGTGCTACCACTCGTGTAG
    TTACATAGCCTTTCTCGATAATATTATTTTGTATTTGCTTCATTAAAATGCTCAAACCTTCTCCGCCTAA
    ACAATGCGGTAATCTCAGTTGTAAATCTGCAACCGCTTTATCTAAAGCCCATTGGAATTGCTCGGTTTGA
    CTTAATCTTTCGGACGGATAATCGATTAAGGAAAGATGATGAATGAGGTAGCAGGGAGACTCCGCCGACG
    GAATTTGTAAATTTCTTTCTTGGCGAGTGTCCATTCGGATATCGGTTTGAGGTATAAATTGCGCTTCTTG
    TTGAACGGTTTGCTGTCGATGACGTTGTTGAATTTCACTATCAATTTGAATATTACGCTTTAATTGCGTT
    TCTTTGAAAGGCGGCGGAGCCGTATTTCCCGTATGGGAATATATAAGTAATGCGGAGAATAAATAGAGAT
    GTAAGTAACGCA
  • Protein Sequence : Show Sequence
    >gi|190150285|ref|YP_001968810.1| putative hemolysin activator protein [Actinobacillus pleuropneumoniae serovar 7 str. AP76]
    MRYLHLYLFSALLIYSHTGNTAPPPFKETQLKRNIQIDSEIQQRHRQQTVQQEAQFIPQTDIRMDTRQER
    NLQIPSAESPCYLIHHLSLIDYPSERLSQTEQFQWALDKAVADLQLRLPHCLGGEGLSILMKQIQNNIIE
    KGYVTTRVVAQEQDLGSGNLVLSVIVGKIGNTVVMDNGRVPRFTPLHALTAFTFEKGDILNVRNIEQSLE
    NLK
  • Molecule Role : Protective antigen
  • Molecule Role Annotation : Following a booster vaccination, a significant (P < 0.05) IgG antibody response to the CP, LPS, and HP (APP7_1016) was detected in the vaccinated pigs. The pigs vaccinated with the CP-HP and LPS-HP conjugates exhibited significantly less mortality (P < 0.05) and significantly greater weight gain (P < 0.001) than unvaccinated pigs. Vaccinated pigs exhibited significantly fewer and less extensive gross pulmonary lesions (P < 0.001) when compared with unvaccinated pigs (Byrd et al., 1992).
2. apxIA
  • Gene Name : apxIA
  • Sequence Strain (Species/Organism) : Actinobacillus pleuropneumoniae L20
  • NCBI Gene ID : 4850247
  • NCBI Protein GI : 126208908
  • Locus Tag : APL_1444
  • Genbank Accession : CP000569
  • Protein Accession : YP_001054133
  • Taxonomy ID : 416269
  • Gene Starting Position : 1652926
  • Gene Ending Position : 1655994
  • Gene Strand (Orientation) : -
  • Protein Name : RTX-I toxin determinant A
  • Protein pI : 5.34
  • Protein Weight : 104673.05
  • Protein Length : 1022
  • Protein Note : pubmed:8359891
  • DNA Sequence : Show Sequence
    >gi|126207488:1652926-1655994 Actinobacillus pleuropneumoniae L20, complete genome
    CTTAAGCAGATTGTGTTAAATAATTACTGAAAGCATTCGGTATGCTATTCGGTGAATTGCTGTAAGCTAA
    TGGACCGCCTGCTCCCACTAAAACTTGTCCGTGTTCACCGGTAAGTGCACCAGTAGATGAAATGATTTGT
    TCAATATCTTTTCTGAGTTGAGGAAGTGCATTATATCCTTCTCCATAATACAATTTTTCTTCTAACCCAT
    TAAAGTAATTGATACCTCTTCCAGGAGCTGTCAGTTCAGTCAGATATTGGCTTAAAACATAAGCTCCATC
    TTTATCAACGATTTTTTCAATTTTATTATTTTGTCCTTCTTTACCTTCCTTGAACCAATCTTTAATCGTG
    AGCGCATTCCCATTGTAATCTTCATGGTAATACAGTGTTCCTCCGATTCTTTTATTCACAAGAAGATCAT
    CACCGATTCTGATAAATCCTACATCTTTAAAACTAAGATCCGATAACAATAACGTATCATCATCACCGCC
    TTTCTCAATAATGATATGACGACCGTACTCCTTACTGTAGCGATAAATATCCTTACCTAAACCACCGTAG
    ATTTTGTCATTGCCTACACCACCGTCAAATAAGTTAGTACCATCGCTGCCATACAGAATGTCATTACCCG
    CACCACCTAATAATACGTTGTATTGACCCTCAAAGACCTGCAACTCATCGTCACCATCACCGCCATTAAG
    GAAATTATTACCTTTTCCGCCGATTAATCGGTCGTTTCCGTTACCACCAACAAGATGGTCGTTACCGTCA
    CCGCCATGGATTACATCATTACCGTCGTCTCCGTATAAGATATCGTGGCCGTCATTACCGTAGATTTCAT
    CATCGCCTTTCGCACCATGGAAAATATCGGTAAAGCGACTACCAAAGAATTTGTCTTTACGATTACTACC
    GATAATTTCTTCAACAGAATGTAATTCATCTTTAGCTCTGATACCGTTCCCAAGTTCGAATGGGCTTAAC
    TCATAATCACGATATTCTAATTTTTCACTGCGTTTTCCAACTGAAATATCCTGAGTTTTAACCACTTCTT
    TTAAAACTTTTACATCAGCTTTAAGTTCTTTAGTGACAATATATTCACCGGCTTTCTGTGCACTTTGTCC
    GTCAAATGTTAAGTAACCTGTATCGGTTTTATCGTAATATGCTACATCATGTCCGTTACCCGCATATACG
    ATAGATGAACCGGATGAAAGATATATACGATCATTTTTCTCACCCAAATGAGATTCAATGGTCACTTGAC
    GTTTTTCACCTTTTTTATCTATCGCTAATTGGATAAGATTCGTATAGTCATAAATCGCATTATGTGACTG
    CACACCGGTTACCACCCATTTTTCTTTACCTTTAACGAATAATTCGGTCATATATTCGTATTTACCGGTT
    TGCTTACGCTCACGAATCTCTTCACCTGCGGTAAAAACCGGTGTAATAAATTTCAATAAAGTGGTTTTGT
    TAATTGAAGAAAGGTCGATTTTGCCTTCAAGCGGATCAAACACTTTTTTATCAAAACGATCCGGATCTTT
    CTCTAACAATTTTCCTTCTTCAAAGAAATCGACATAAGCCTTACCGCTTTTCGCATCCGCACCTTTACGC
    GTGATACCGGCAAGTTCCCCGATATTGACATCCCAACGTTGTTGCGTAATAGCAACGACACGCTCTACCG
    AATACTCTTTATTGTATTGTGATAACAATTCAAAGGTATCTTCTAAGAATGCGGAATGGCGGGCGTCATA
    ACCGTTTTCAAAATAGTTTTTACCGTGTTTTTTCTCCCATTCGTCAATCTTATTCGCTAATTTCGTTGCA
    ACTCGTTCGAAGATTGCCTGTTTAGAAGCATCTAAAATACCTGAAATAATACCGGTGATTGCACTAACTA
    AAGCTGCTACCGGCGCACCGACTAATGAGCCGGTTGCAGCAGCCCCAACACCTGCGGAAGCCGCACTTAA
    CACACTGTTAATCGTGGTTAATGCCGCTTCAATCGCACCGGTTTCACGGTAGAATGAAGCTAATAAACTA
    TCACCTTCATAACCGAACTTTTTAAAGCGCTCCGAATATTGTTCAAGCTGTTTCGCACGTTCAAACTTAT
    CCGCAACATTTAAGAACGAAAGCGGGCTAATCGCTAATGCTACGACCGAACCGATTAAACCACCGGTTGC
    CGCAGTTGTGGATAAGCCTGCCGCCACACGTTGCGCAATAATATATTGAGAAACCGCTTTACCGATATTG
    CCTAAGATTTTAGTTGAGATTTCAATACCTGCCGCCGCTTTTGTACCTGCATCGGCATCTTTATTACTTA
    AAATGAATGAAGCCGAAACAACAGATAAGATACCTGATACGGCATCAAACCCAGGTCCTGCAAGGCTTAA
    ATCTGGAAGGTTATTTAACTTACTTGCTAAACCGCTTAAGCGAGTGTTAGATAAGGCATTGCCCAATTTA
    CCTAATTGTTCGGCAAACGCATCCACCGTACCCGTTGCACTTGCAATGTTATCCACTAACTGAGCGGCTA
    GATCCACACCCGCTTTAGCTAATTCCGAACCACTGACGTCCTCACCGTTTCTACGGCGACGAAGCAGGCT
    ATCAAGATCCATACCCGCAATAGCCGTACCTAAGAAACTTTGTAAGGCTGAAAGTGCCGTTTGCGTTTTA
    TTTAAACGTTGTCCTAATGCTTCCGATGAACCGCCGAGCGATTTACTTAATTTTTGGTTCTTATTCAGTA
    ACTTATCAAACTGAGGTGCAAATAATGCGATGCCTCGTTCCGAGAAACCTAATAGTTTTTCCGTTGTACC
    GAATAATTCTTTCGCTAGTGCGGTACCGTTTTTTTCCGAGCGATGTACTTCGATCCCTAAAGCCTCCGCC
    GCTTTCACTAAATCATTAAGACTTGAGCCGGTACTAGCTTGATAATCTTTCGGAATATATAAAATTAATT
    TCTGCCCTGCTTGCTTCACCTGTCCCAAACCATTTTTTAATGCGCCGGCACCTGATTTAGCTGCACTTTT
    TGTATGTTGATTAAGTGAATCAATCAATCCTTTGACTCTATCGAGCTGAGAGTTAGCCA
  • Protein Sequence : Show Sequence
    >gi|126208908|ref|YP_001054133.1| RTX-I toxin determinant A [Actinobacillus pleuropneumoniae L20]
    MANSQLDRVKGLIDSLNQHTKSAAKSGAGALKNGLGQVKQAGQKLILYIPKDYQASTGSSLNDLVKAAEA
    LGIEVHRSEKNGTALAKELFGTTEKLLGFSERGIALFAPQFDKLLNKNQKLSKSLGGSSEALGQRLNKTQ
    TALSALQSFLGTAIAGMDLDSLLRRRRNGEDVSGSELAKAGVDLAAQLVDNIASATGTVDAFAEQLGKLG
    NALSNTRLSGLASKLNNLPDLSLAGPGFDAVSGILSVVSASFILSNKDADAGTKAAAGIEISTKILGNIG
    KAVSQYIIAQRVAAGLSTTAATGGLIGSVVALAISPLSFLNVADKFERAKQLEQYSERFKKFGYEGDSLL
    ASFYRETGAIEAALTTINSVLSAASAGVGAAATGSLVGAPVAALVSAITGIISGILDASKQAIFERVATK
    LANKIDEWEKKHGKNYFENGYDARHSAFLEDTFELLSQYNKEYSVERVVAITQQRWDVNIGELAGITRKG
    ADAKSGKAYVDFFEEGKLLEKDPDRFDKKVFDPLEGKIDLSSINKTTLLKFITPVFTAGEEIRERKQTGK
    YEYMTELFVKGKEKWVVTGVQSHNAIYDYTNLIQLAIDKKGEKRQVTIESHLGEKNDRIYLSSGSSIVYA
    GNGHDVAYYDKTDTGYLTFDGQSAQKAGEYIVTKELKADVKVLKEVVKTQDISVGKRSEKLEYRDYELSP
    FELGNGIRAKDELHSVEEIIGSNRKDKFFGSRFTDIFHGAKGDDEIYGNDGHDILYGDDGNDVIHGGDGN
    DHLVGGNGNDRLIGGKGNNFLNGGDGDDELQVFEGQYNVLLGGAGNDILYGSDGTNLFDGGVGNDKIYGG
    LGKDIYRYSKEYGRHIIIEKGGDDDTLLLSDLSFKDVGFIRIGDDLLVNKRIGGTLYYHEDYNGNALTIK
    DWFKEGKEGQNNKIEKIVDKDGAYVLSQYLTELTAPGRGINYFNGLEEKLYYGEGYNALPQLRKDIEQII
    SSTGALTGEHGQVLVGAGGPLAYSNSPNSIPNAFSNYLTQSA
  • Molecule Role : Virmugen
  • Molecule Role Annotation : The mutant strain, with deletion of apxIA gene, obtained in this study offered a level of cross-serovar protection against A. pleuropneumoniae infection in mice and pigs (Xu et al., 2006).
  • Related Vaccine(s): Actinobacillus pleuropneumoniae apxIA mutant vaccine
3. ApxIA
  • Gene Name : ApxIA
  • Sequence Strain (Species/Organism) : Actinobacillus pleuropneumoniae
  • VO ID : VO_0011019
  • NCBI Protein GI : 312899
  • Other Database IDs : CDD:111291
    CDD:164328
    CDD:116920
    GOA:P55128
    InterPro: IPR001343
    InterPro: IPR003995
    InterPro: IPR011049
    InterPro: IPR013550
    UniProtKB/UniProt: P55128
  • Taxonomy ID : 715
  • Gene Strand (Orientation) : ?
  • Protein pI : 5.34
  • Protein Weight : 104673.05
  • Protein Length : 1022
  • Protein Note : RTX N-terminal domain; pfam02382
  • Protein Sequence : Show Sequence
    >gi|312899|emb|CAA51548.1| apxIA [Actinobacillus pleuropneumoniae]
    MANSQLDRVKGLIDSLNQHTKSAAKSGAGALKNGLGQVKQAGQKLILYIPKDYQASTGSSLNDLVKAAEA
    LGIEVHRSEKNGTALAKELFGTTEKLLGFSERGIALFAPQFDKLLNKNQKLSKSLGGSSEALGQRLNKTQ
    TALSALQSFLGTAIAGMDLDSLLRRRRNGEDVSGSELAKAGVDLAAQLVDNIASATGTVDAFAEQLGKLG
    NALSNTRLSGLASKLNNLPDLSLAGPGFDAVSGILSVVSASFILSNKDADAGTKAAAGIEISTKILGNIG
    KAVSQYIIAQRVAAGLSTTAATGGLIGSVVALAISPLSFLNVADKFERAKQLEQYSERFKKFGYEGDSLL
    ASFYRETGAIEAALTTINSVLSAASAGVGAAATGSLVGAPVAALVSAITGIISGILDASKQAIFERVATK
    LANKIDEWEKKHGKNYFENGYDARHSAFLEDTFELLSQYNKEYSVERVVAITQQRWDVNIGELAGITRKG
    SDTKSGKAYVDFFEEGKLLEKEPDRFDKKVFDPLEGKIDLSSINKTTLLKFVTPVFTAGEEIRERKQTGK
    YEYMTELFVKGKEKWVVTGVQSHNAIYDYTNLIQLAIDKKGEKRQVTIESHLGEKNDRIYLSSGSSIVYA
    GNGHDVAYYDKTDTGYLTFDGQSAQKAGEYIVTKELKADVKVLKEVVKTQDISVGKRSEKLEYRDYELSP
    FELGNGIRAKDELHSVEEIIGSNRKDKFFGSRFTDIFHGAKGDDEIYGNDGHDILYGDDGNDVIHGGDGN
    DHLVGGNGNDRLIGGKGNNFLNGGDGDDELQVFEGQYNVLLGGAGNDILYGSDGTNLFDGGVGNDKIYGG
    LGKDIYRYSKEYGRHIIIEKGGDDDTLLLSDLSFKDVGFIRIGDDLLVNKRIGGTLYYHEDYNGNALTIK
    DWFKEGKEGQNNKIEKIVDKDGAYVLSQYLTELTAPGRGINYFNGLEEKLYYGEGYNALPQLRKDIEQII
    SSTGAFTGDHGKVSVGSGGPLVYNNSANNVANSLSYSLAQAA
  • Molecule Role : Protective antigen
  • Molecule Role Annotation : The ApxI antigen is an important factor in the protection against Actinobacillus pleuropneumoniae serotype 5 infection; therefore, the protective immunity in mice following oral immunization with Saccharomyces cerevisiae expressing either ApxIA (group C) or ApxIIA (group D) alone or both (group E) was compared with that in two control groups (group A and B). The immunogenicity of the rApxIA antigen derived from the yeast was confirmed by a high survival rate and an ApxIA-specific IgG antibody response (p < 0.01) (Shin et al., 2007).
  • Related Vaccine(s): A. pleuropneumoniae ApxIa and ApxIIa protein vaccine , A. pleuropneumoniae ApxIA protein vaccine , HS93Tox-/pIG-T1K , rS.C-APP-ApxI/ApxII
4. apxIA
  • Gene Name : apxIA
  • Sequence Strain (Species/Organism) : Actinobacillus pleuropneumoniae strain CH80-1 serotype 1
  • NCBI Nucleotide GI : 18026876
  • NCBI Protein GI : 18026877
  • Protein Accession : AAL55666.1
  • Taxonomy ID : 715
  • Gene Strand (Orientation) : ?
  • Protein Name : ApxIA
  • Protein Note : isolated from an infected pig
  • DNA Sequence : Show Sequence
    >gi|18026876|gb|AF240779.1| Actinobacillus pleuropneumoniae ApxIA (apxIA) gene, complete cds
    ATGGCTAACTCTCAGCTCGATAGAGTCAAAGGATTGATTGATTCACTTAATCAACATACAAAAAGTGCAG
    CTAAATCAGGTGCCGGCGCATTAAAAAATGGTTTGGGACAGGTGAAGCAAGCAGGGCAGAAATTAATTTT
    ATATATTCCGAAAGATTATCAAGCTAGTACCGGCTCAAGTCTTAATGATTTAGTGAAAGCGGCGGAGGCT
    TTAGGGATCGAAGTACATCGCTCGGAAAAAAACGGTACCGCACTAGCGAAAGAATTATTCGGTACAACGG
    AAAAACTATTAGGTTTCTCGGAACGAGGCATCGCATTATTTGCACCTCAGTTTGATAAGTTACTGAATAA
    GAACCAAAAATTAAGTAAATCGCTCGGCGGCTCATCGGAAGCATTAGGACAACGTTTAAATAAAACGCAA
    ACGGCACTTTCAGCCTTACAAAGTTTCTTAGGTACGGCTATTGCGGGTATGGATCTTGATAGCCTGCTTC
    GTCGCCGTAGAAACGGTGAGGACGTCAGTGGTTCGGAATTAGCTAAAGCAGGTGTGGATCTAGCCGCTCA
    GTTAGTGGATAACATTGCAAGTGCAACGGGTACGGTGGATGCGTTTGCCGAACAATTAGGTAAATTGGGC
    AATGCCTTATCTAACACTCGCTTAAGCGGTTTAGCAAGTAAGTTAAATAACCTTCCAGATTTAAGCCTTG
    CAGGACCTGGGTTTGATGCCGTATCAGGTATCTTATCTGTTGTTTCGGCTTCATTCATTTTAAGTAATAA
    AGATGCCGATGCAGGTACAAAAGCGGCGGCAGGTATTGAAATCTCAACTAAAATCTTAGGCAATATCGGT
    AAAGCGGTTTCTCAATATATTATTGCGCAACGTGTGGCGGCAGGCTTATCCACAACTGCGGCAACCGGTG
    GTTTAATCGGTTCGGTCGTAGCATTAGCGATTAGCCCGCTTTCGTTCTTAAATGTTGCGGATAAGTTTGA
    ACGTGCGAAACAGCTTGAACAATATTCGGAGCGCTTTAAAAAGTTCGGTTATGAAGGTGATAGTTTATTA
    GCTTCATTCTACCGTGAAACCGGTGCGATTGAAGCGGCATTAACCACGATTAACAGTGTGTTAAGTGCGG
    CTTCCGCAGGTGTTGGGGCTGCTGCAACCGGCTCATTAGTCGGTGCGCCGGTAGCAGCTTTAGTTAGTGC
    AATCACCGGTATTATTTCAGGTATTTTAGATGCTTCTAAACAGGCAATCTTCGAACGAGTTGCAACGAAA
    TTAGCGAATAAGATTGACGAATGGGAGAAAAAACACGGTAAAAACTATTTTGAAAACGGTTATGACGCCC
    GCCATTCCGCATTCTTAGAAGATACCTTTGAATTGTTATCACAATACAATAAAGAGTATTCGGTAGAGCG
    TGTCGTTGCTATTACGCAACAGCGTTGGGATGTCAATATCGGTGAACTTGCCGGCATTACTCGCAAAGGT
    TCTGATACGAAAAGCGGTAAAGCTTACGTTGATTTCTTTGAAGAAGGAAAACTTTTAGAGAAAGAACCGG
    ATCGTTTTGATAAAAAAGTGTTTGATCCGCTTGAAGGTAAAATCGACCTTTCTTCAATTAACAAAACCAC
    TTTATTGAAATTTGTTACGCCGGTCTTTACCGCAGGTGAAGAGATTCGTGAGCGTAAGCAAACCGGTAAA
    TACGAATATATGACCGAATTATTCGTTAAAGGTAAAGAAAAATGGGTGGTAACCGGTGTGCAGTCACATA
    ATGCGATTTATGACTATACGAATCTTATCCAATTAGCGATAGATAAAAAAGGTGAAAAACGTCAAGTGAC
    CATTGAATCTCATTTGGGTGAGAAAAATGATCGTATATATCTTTCATCCGGTTCATCTATCGTATATGCG
    GGTAACGGACATGATGTAGCATATTACGATAAAACCGATACAGGTTACTTAACATTTGACGGACAAAGTG
    CACAGAAAGCCGGTGAATATATTGTCACTAAAGAACTTAAAGCTGATGTAAAAGTTTTAAAAGAAGTGGT
    TAAAACTCAGGATATTTCAGTTGGAAAACGCAGTGAAAAATTAGAATATCGTGATTATGAGTTAAGCCCA
    TTCGAACTTGGGAACGGTATCAGAGCTAAAGATGAATTACATTCTGTTGAAGAAATTATCGGTAGTAATC
    GTAAAGACAAATTCTTTGGTAGTCGCTTTACCGATATTTTCCATGGTGCGAAAGGCGATGATGAAATCTA
    CGGTAATGACGGCCACGATATCTTATACGGAGACGACGGTAATGATGTAATCCATGGCGGTGACGGTAAC
    GACCATCTTGTTGGTGGTAACGGAAACGACCGATTAATCGGCGGAAAAGGTAATAATTTCCTTAATGGCG
    GTGATGGTGACGATGAGTTGCAGGTCTTTGAGGGTCAATACAACGTATTATTAGGTGGTGCGGGTAATGA
    CATTCTGTATGGCAGCGATGGTACTAACTTATTTGACGGTGGTGTAGGCAATGACAAAATCTACGGTGGT
    TTAGGTAAGGATATTTATCGCTACAGTAAGGAGTACGGTCGTCATATCATTATTGAGAAAGGCGGTGATG
    ATGATACGTTATTGTTATCGGATCTTAGTTTTAAAGATGTAGGATTTATCAGAATCGGTGATGATCTTCT
    TGTGAATAAAAGAATCGGAGGAACACTGTATTACCATGAAGATTACAATGGGAATGCGCTCACGATTAAA
    GATTGGTTCAAGGAAGGTAAAGAAGGACAAAATAATAAAATTGAAAAAATCGTTGATAAAGATGGAGCTT
    ATGTTTTAAGCCAATATCTGACTGAACTGACAGCTCCTGGAAGAGGTATCAATTACTTTAATGGGTTAGA
    AGAAAAATTGTATTATGGAGAAGGATATAATGCACTTCCTCAACTCAGAAAAGATATTGAACAAATCATT
    TCATCTACGGGTGCATTTACCGGTGATCACGGAAAAGTATCTGTAGGCTCAGGCGGACCGTTAGTCTATA
    ATAACTCAGCTAACAATGTAGCAAATTCTTTGAGTTATTCTTTAGCACAAGCAGCTTAA
  • Molecule Role : Protective antigen
  • Related Vaccine(s): A. pleuropneumoniae DNA vaccine pcDNA-apxIA/pcDNA-apxIIA
5. ApxIC
  • Gene Name : ApxIC
  • Sequence Strain (Species/Organism) : Actinobacillus pleuropneumoniae
  • NCBI Protein GI : 1477456
  • Other Database IDs : CDD:154216
  • Taxonomy ID : 715
  • Gene Strand (Orientation) : ?
  • Protein Length : 172
  • Protein Note : RTX toxin acyltransferase family; cl01131
  • Protein Sequence : Show Sequence
    >gi|1477456|gb|AAB17219.1| apxIC [Actinobacillus pleuropneumoniae]
    MSKKINGFEVLGEVAWLWASSPLHRKWPLSLLAINVLPAIESNQYVLLKRDGFPIAFCSWANLNLENEIK
    YLDDVASLVADDWTSGDRRWFIDWIAPFGDSAALYKHMRDNFPNELFRAIRVDPDSRVGKISEFHGGKID
    KKLASKIFQQYHFELMSELKNKQNFKFSLVNS
  • Molecule Role : Virmugen
  • Molecule Role Annotation : In the intra-peritoneal mouse model, the virulence of the mutant strain decreased at least 100 fold compared with the parent strain. The mutant was evaluated as a potential vaccine using a vaccination-challenge trial. The death number and lung lesion score in the vaccinated pigs given the serovar 1 challenge were obviously lower than those in the unvaccinated pigs (Xu et al., 2007).
  • Related Vaccine(s): Actinobacillus pleuropneumoniae ApxIC mutant vaccine
6. ApxIIA
  • Gene Name : ApxIIA
  • Sequence Strain (Species/Organism) : Actinobacillus pleuropneumoniae
  • VO ID : VO_0011020
  • NCBI Protein GI : 52630374
  • Other Database IDs : CDD:111291
    CDD:116920
  • Taxonomy ID : 715
  • Gene Strand (Orientation) : ?
  • Protein Name : ApxIIA
  • Protein pI : 5.34
  • Protein Weight : 96054.51
  • Protein Length : 956
  • Protein Note : biovar: 1
  • Protein Sequence : Show Sequence
    >gi|52630374|gb|AAU84700.1| ApxIIA [Actinobacillus pleuropneumoniae]
    MSKITLSSLKSSLQQGLKNGKNKLNQAGTTLKNGLTQTGHSLQNGAKKLILYIPQGYDSGQGNGVQDLVK
    AANDLGIEVWREERSNLDIAKTSFDTTQKILGFTDRGIVLFAPQLDNLLKKNPKIGNTLGSASSISQNIG
    KANTVLGGIQSILGSVLSGVNLNELLQNKDPNQLELAKAGLELTNELVGNIASSVQTVDAFAEQISKLGS
    HLQNVKGLGGLSNKLQNLPDLGKASLGLDIISGLLSGASAGLILADKEASTEKKAAAGVEFANQIIGNVT
    KAVSSYILAQRVASGLSSTGPVAALIASTVALAVTPLSFLNVADKFKQADLIKSYSERFQKLGYDGDRLL
    ADFHRETGTIDASVTTINTALAAISGGVGAASAGSLVGAPVALLVAGVTGLITTILEYSKQAMFEHVANK
    VHDRIVEWEKKHNKNYFEQGYDSRHLADLQDNMKFLINLNKELQAERVVAITQQRWDNQIGDLAAISRRT
    DKISSGKAYVDAFEGGQHQSYDSSVQLDNKNGIINISNTNRKTQSVLFRTPLLTPGEENRERIQEGKNSY
    ITKLHIQRVDSWTVTDGDASSSVDFTNVVQRIAVKFDDAGNIIESKDTKIIANLGAGNDNVFVGSSTTVI
    DGGDGHDRVHYSRGEYGALVIDATAETEKGSYSVKRYVGDSKALHETIATHQTNVGNREEKIEYRREDDR
    FHTGYTVTDSLKSVEEIIGSQFNDIFKGSQFDDVFHGGNGVDTIDGNDGDDHLFGGAGDDVIDGGNGNNF
    LVGGTGNDIISGGKDNDIYVHKTGDGNDSITDSGGQDKLAFSDVNLKDLTFKKVDSSLEIINQKGEKVRI
    GNWFLEDDLASTVANYKATNDRKIEEIIGKGGERITSEQVDKLIKEGNNQISAEALSKVVNDYNTSKDRQ
    NVSNSLAKLISPVGSFTSSSDFRNNLGTYVPSSIDVSNNIQLARAA
  • Molecule Role : Protective antigen
  • Molecule Role Annotation : The levels of local and systemic antibodies specific to the Apx antigens were investigated in mice orally immunized with Apx antigen-expressing yeast. Specific IgA responses to ApxIA or ApxIIA in the intestines and lungs from mice immunized with yeast expressing Apx antigens were significantly higher than those in the control groups after the second and third immunizations, respectively (p < 0.05). All mice were monitored for up to 72 h after challenge with the MLD of an A. pleuropneumoniae field isolate. Overall, the final survival rates of the vaccinated groups were higher than those of the control groups at each time point (Shin et al., 2007).
  • Related Vaccine(s): A. pleuropneumoniae ApxIa and ApxIIa protein vaccine , rS.C-APP-ApxI/ApxII
7. apxIIA
  • Gene Name : apxIIA
  • Sequence Strain (Species/Organism) : Actinobacillus pleuropneumoniae serotype 2
  • NCBI Protein GI : 255959000
  • Locus Tag : APL_0956
  • Genbank Accession : CP000569
  • Protein Accession : YP_001053657
  • Other Database IDs : CDD:111291
    CDD:116920
  • Taxonomy ID : 715
  • Gene Starting Position : 1095617
  • Gene Ending Position : 1098487
  • Gene Strand (Orientation) : -
  • Protein Name : RTX-II toxin determinant A
  • Protein pI : 5.34
  • Protein Weight : 96054.51
  • Protein Length : 956
  • Protein Note : RTX N-terminal domain; pfam02382
  • DNA Sequence : Show Sequence
    >gi|126207488:1095617-1098487 Actinobacillus pleuropneumoniae L20, complete genome
    ATTAAGCGGCTCTAGCTAATTGAATATTATTCGAGACATCTATTGATGAAGGAACATATGTTCCTAAATT
    ATTCCTAAAGTCTGAGGAAGACGTAAAGCTCCCGACTGAAGAAATCAATTTTGCTAAGCTATTAGATACG
    TTCTGTCTATCTTTACTCGTATTGTAATCATTCACAACTTTGGATAATGCTTCTGCAGAGATTTGATTGT
    TACCCTCCTTAATCAGTTTATCAACTTGTTCTGATGTAATACGTTCTCCTCCTTTACCAATAATTTCCTC
    AATTTTTCGGTCATTCGTAGCTTTATAGTTAGCAACTGTGCTAGCCAAATCATCTTCTAAGAACCAATTC
    CCAATACGAACTTTTTCTCCTTTTTGATTAATGATTTCGAGAGAAGAATCTACTTTCTTAAAGGTGAGGT
    CTTTAAGATTTACATCCGAAAATGCCAGTGTATCTTGTCCGCCAGAGTCTGTAATAGAATCATTTCCATC
    GCCTGTTTTATGGACATATATATCATTATCTTTACCTCCCGAGATAATATCATTACCGGTTCCTCCAACA
    AGGAAATTGTTACCGTTTCCTCCATCGATAACATCATCGCCTGCGCCACCAAATAAATGATCGTCACCAT
    CGTTACCATCAATAGTGTCTACACCATTACCACCATGGAACACATCATCAAATTGGCTTCCTTTGAAAAT
    ATCATTAAATTGTGAACCAATGATCTCTTCAACTGATTTGAGTGAGTCCGTCACAGTATAACCAGTATGA
    AAACGATCATCTTCACGACGATATTCAATTTTTTCTTCACGATTACCAACATTTGTTTGGTGGGTGGCAA
    TTGTTTCATGTAATGCTTTACTGTCTCCGACATAGCGTTTTACTGAATATGAGCCTTTTTCTGTCTCGGC
    TGTAGCATCAATAACTAATGCGCCATATTCTCCTCTACTGTAGTGAACTCGATCATGTCCGTCCCCGCCA
    TCAATAACGGTAGTACTTGACCCAACAAATACATTATCGTTACCAGCACCTAAATTTGCGATAATTTTAG
    TATCTTTAGATTCGATAATGTTACCTGCATCATCAAATTTCACAGCGATTCGTTGTACTACATTAGTGAA
    ATCTACGCTTGAGCTAGCATCACCATCTGTTACAGTCCAACTGTCAACTCTTTGTATATGTAATTTTGTA
    ATATAAGAATTTTTACCTTCCTGAATACGTTCCCGATTCTCTTCACCTGGAGTTAGTAATGGAGTTCTGA
    ATAAAACACTTTGTGTCTTTCTATTTGTATTACTAATATTAATAATACCGTTTTTGTTATCTAGCTGTAC
    GGATGAATCGTAGGACTGGTGTTGCCCCTCCTCAAAAGCATCCACATAAGCTTTTCCACTGGAAATTTTA
    TCCGTTCTACGGCTAATTGCCGCTAGGTCTCCAATTTGGTTATCCCATCTTTGTTGGGTAATAGCTACTA
    CGCGTTCAGCCTGAAGTTCTTTATTTAAATTGATAAGAAACTTCATATTGTCTTGTAAATCAGCTAAATG
    ACGAGAATCATAACCTTGCTCAAAATAGTTTTTATTATGTTTTTTCTCCCATTCAACTATTCTGTCATGA
    ACCTTATTTGCAACATGTTCAAACATGGCTTGTTTAGAATATTCTAGAATAGTTGTAATAAGTCCCGTAA
    CACCAGCAACGAGTAACGCAACTGGAGCTCCGACTAGAGAACCCGCGCTTGCAGCTCCAACTCCACCGGA
    GATAGCTGCTAAAGCAGTGTTAATTGTTGTTACAGAAGCATCAATAGTTCCTGTCTCACGGTGAAAATCA
    GCTAATAAACGATCTCCATCATATCCTAATTTTTGGAAGCGTTCAGAATATGATTTGATTAAATCAGCTT
    GTTTAAACTTATCAGCTACATTTAAGAATGAAAGAGGGCTAACAGCTAGTGCAACTGTAGATGCGATTAA
    TGCAGCGACAGGACCAGTTGAAGACAAACCTGAAGCGACTCGTTGGGCAAGAATGTAAGATGAGACCGCT
    TTTGTTACATTACCTATAATTTGGTTAGCAAATTCTACACCTGCGGCAGCTTTCTTTTCTGTTGAAGCCT
    CTTTATCTGCTAAAATGAGACCTGCAGATGCTCCAGAAAGTAAACCAGAGATAATGTCCAAACCTAAACT
    TGCTTTTCCTAGATCTGGTAGATTTTGTAATTTATTACTCAATCCTCCTAATCCTTTCACATTCTGTAAA
    TGTGAACCTAGTTTAGATATTTGTTCTGCAAATGCATCTACAGTTTGCACCGAGCTAGCAATATTACCAA
    CTAATTCATTAGTCAGTTCTAGCCCTGCTTTTGCAAGTTCTAATTGATTAGGATCTTTATTTTGAAGTAA
    TTCATTCAGATTTACTCCAGATAAAACAGATCCTAAAATAGATTGAATACCACCTAATACAGTATTGGCT
    TTACCTATATTTTGTGAGATGCTAGAAGCACTTCCTAATGTATTGCCAATTTTAGGATTCTTCTTTAATA
    AATTATCTAGCTGAGGTGCAAATAATACAATTCCTCTATCAGTAAAACCTAGAATTTTCTGAGTTGTATC
    AAAGCTAGTTTTTGCAATGTCCAAATTGCTGCGTTCTTCTCGCCATACTTCAATACCTAAATCATTAGCA
    GCTTTAACTAAATCTTGAACTCCATTTCCTTGACCCGAATCATAGCCTTGAGGAATATATAAGATTAATT
    TTTTAGCCCCATTCTGTAGAGAATGACCAGTTTGAGTTAAACCATTCTTCAGTGTTGTACCTGCTTGATT
    TAACTTGTTTTTCCCATTTTTCAATCCTTGTTGTAAGGACGATTTTAATGATGACAAAGTGATTTTTGAC
    A
  • Protein Sequence : Show Sequence
    >gi|255959000|gb|ACU43588.1| RTX-II toxin determinant A [Actinobacillus pleuropneumoniae]
    MSKITLSSLKSSLQQGLKNGKNKLNQAGTTLKNGLTQTGHSLQNGAKKLILYIPQGYDSGQGNGVQDLVK
    AANDLGIEVWREERSNLDIAKTSFDTTQKILGFTDRGIALFAPQLDNLLKKNPKIGNTLGSASSISQNIG
    KANTVLGGIQSILGSVLSGVNLNELLQNKDPNQLELAKAGLELTNELVGNIASSVQTVDAFAEQISKLGS
    HLQNVKGLGGLSNKLQNLPDLGKASLGLDIISGLLSGASAGLILADKEASTEKKAAAGVEFANQIIGNVT
    KAVSSYILAQRVASGLSSTGPVAALIASTVALAVSPLSFLNVADKFKQADLIKSYSERFQKLGYDGDRLL
    ADFHRETGTIDASVTTINTALAAISGGVGAASAGSLVGAPVALLVAGVTGLITTILEYSKQAMFEHVANK
    VHDRIVEWEKKHNKNYFEQGYDSRHLADLQDNMKFLINLNKELQAERVVAITQQRWDNQIGDLAAISRRT
    DKISSGKAYVDAFEEGQHQSYDSSVQLDNKNGIINISNTNRKTQSVLFRTPLLTPGEENRERIQEGKNSY
    ITKLHIQRVDSWTVTDGDASSSVDFTNVVQRIAVKFDDAGNIIESKDTKIIANLGAGNDNVFVGSSTTVI
    DGGDGHDRVHYSRGEYGALVIDATAETEKGSYSVKRYVGDSKALHETIATHQTNVGNREEKIEYRREDDR
    FHTGYTVTDSLKSVEEIIGSQFNDIFKGSQFDDVFHGGNGVDTIDGNDGDDHLFGGAGDDVIDGGNGNNF
    LVGGTGNDIISGGKDNDIYVHKTGDGNDSITDSGGQDKLAFSDVNLKDLTFKKVDSSLEIINQKGEKVRI
    GNWFLEDDLASTVANYKATNDRKIEEIIGKGGERITSEQVDKLIKEGNNQISAEALSKVVNDYNTSKDRQ
    NVSNSLAKLISSVGSFTSSSDFRNNLGTYVPSSIDVSNNIQLARAA
  • Molecule Role : Virmugen
  • Molecule Role Annotation : Deletion in ureC and apxIIA genes of Actinobacillus pleuropneumoniae serotype 2 prototype provides attenuation and protection in pigs against challenge with the virulent parent strain of A. pleuropneumoniae (Tonpitak et al., 2002).
  • Related Vaccine(s): Actinobacillus pleuropneumoniae ureC and apxIIA double mutant deletion vaccine , rS.C-APP-ApxIIA
8. apxIIA
  • Gene Name : apxIIA
  • Sequence Strain (Species/Organism) : Actinobacillus pleuropneumoniae
  • NCBI Nucleotide GI : 13937106
  • NCBI Protein GI : 13937107
  • Protein Accession : AAK50052.1
  • Taxonomy ID : 715
  • Gene Strand (Orientation) : ?
  • Protein Name : RTX toxin IIA
  • Protein Note : ApxIIA
  • DNA Sequence : Show Sequence
    >gi|13937106|gb|AF363362.1| Actinobacillus pleuropneumoniae RTX toxin IIA gene, complete cds
    ATGTCAAAAATCACTTTGTCATCATTAAAATCGTCCTTACAACAAGGATTGAAAAATGGGAAAAACAAGT
    TAAATCAAGCAGGTACAACACTGAAGAATGGTTTAACTCAAACTGGTCATTCTCTACAGAATGGGGCTAA
    AAAATTAATCTTATATATTCCTCAAGGCTATGATTCGGGTCAAGGAAATGGAGTTCAAGATTTAGTTAAA
    GCTGCTAATGATTTAGGTATTGAAGTATGGCGAGAAGAACGCAGCAATTTGGACATTGCAAAAACTAGCT
    TTGATACAACTCAGAAAATTCTAGGTTTTACTGATAGAGGAATTGTATTATTTGCACCTCAGCTAGATAA
    TTTATTAAAGAAGAATCCTAAAATTGGCAATACATTAGGAAGTGCTTCTAGCATCTCACAAAATATAGGT
    AAAGCCAATACTGTATTAGGTGGTATTCAATCTATTTTAGGATCTGTTTTATCTGGAGTAAATCTGAATG
    AATTACTTCAAAATAAAGATCCTAATCAATTAGAACTTGCAAAAGCAGGGCTAGAACTGACTAATGAATT
    AGTTGGTAATATTGCTAGCTCGGTGCAAACTGTAGATGCATTTGCAGAACAAATATCTAAACTAGGTTCA
    CATTTACAGAATGTGAAAGGATTAGGAGGATTGAGTAATAAATTACAAAATCTACCAGATCTAGGAAAAG
    CAAGTTTAGGTTTGGACATTATCTCTGGTTTACTTTCTGGAGCATCTGCAGGTCTCATTTTAGCAGATAA
    AGAGGCTTCAACAGAAAAGAAAGCTGCCGCAGGTGTAGAATTTGCTAACCAAATTATAGGTAATGTAACA
    AAAGCGGTCTCATCTTACATTCTTGCCCAACGAGTCGCTTCAGGTTTGTCTTCAACTGGTCCTGTCGCTG
    CATTAATCGCATCTACAGTTGCACTAGCTGTTAGCCCTCTTTCATTCTTAAATGTAGCTGATAAGTTTAA
    ACAAGCTGATTTAATCAAATCATATTCTGAACGCTTCCAAAAATTAGGATATGATGGAGATCGTTTATTA
    GCTGATTTTCACCGTGAGACAGGAACTATTGATGCTTCTGTAACAACAATTAACACTGCTTTAGCAGCTA
    TCTCCGGTGGAGTTGGAGCTGCAAGCGCGGGTTCTCTAGTCGGAGCTCCAGTTGCGTTACTCGTTGCTGG
    TGTTACGGGACTTATTACAACTATTCTAGAATATTCTAAACAAGCCATGTTTGAACATGTTGCAAATAAG
    GTTCATGACAGAATAGTTGAATGGGAGAAAAAACATAATAAAAACTATTTTGAGCAAGGTTATGATTCTC
    GTCATTTAGCTGATTTACAAGACAATATGAAGTTTCTTATCAATTTAAATAAAGAACTTCAGGCTGAACG
    CGTAGTAGCTATTACCCAACAAAGATGGGATAACCAAATTGGAGACCTAGCGGCAATTAGCCGTAGAACG
    GATAAAATTTCCAGTGGAAAAGCTTATGTGGATGCTTTTGAGGAGGGGCAACACCAGTCCTACGATTCAT
    CCGTACAGCTAGATAACAAAAACGGTATTATTAATATTAGTAATACAAATAGAAAGACACAAAGTGTTTT
    ATTCAGAACTCCATTACTAACTCCAGGTGAAGAGAATCGGGAACGTATTCAGGAAGGTAAAAATTCTTAT
    ATTACAAAATTACATATACAAAGAGTTGACAGTTGGACTGTAACAGATGGTGATGCTAGCTCAAGCGTAG
    ATTTCACTAATGTAGTACAACGAATCGCTGTGAAATTTGATGATGCAGGTAACATTATAGAATCTAAAGA
    TACTAAAATTATCGCAAATTTAGGTGCTGGTAACGATAATGTATTTGTTGGGTCAAGTACTACCGTTATT
    GATGGCGGGGACGGACATGATCGAGTTCACTACAGTAGAGGAGAATATGGCGCATTAGTTATTGATGCTA
    CAGCCGAGACAGAAAAAGGCTCATATTCAGTAAAACGCTATGTCGGAGACAGTAAAGCATTACATGAAAC
    AATTGCCACCCACCCAACAAATGTTGGTAATCGTGAAGAAAAAATTGAATATCGTCGTGAAGATGATCGT
    TTTCATACTGGTTATACTGTGACGGACTCACTCAAATCAGTTGAAGAGATCATTGGTTCACAATTTAATG
    ATATTTTCAAAGGAAGCCAATTTGATGATGTGTTCCATGGTGGTAATGGTGTAGACACTATTGATGGTAA
    CGATGGTGACGATCATTTATTTGGTGGCGCAGGCGATGATGTTATCGATGGAGGAAACGGTAACAATTTC
    CTTGTTGGAGGAACCGGTAATGATATTATCTCGGGAGGTAAAGATAATGATATTTATGTCCATAAAACAG
    GCGATGGAAATGATTCTATTACAGACTCTGGCGGACAAGATAAACTGGCATTTTCGGATGTAAATCTTAA
    AGACCTCACCTTTAAGAAAGTAGATTCTTCTCTCGAAATCATTAATCAAAAAGGAGAAAAAGTTCGTATT
    GGGAATTGGTTCTTAAAAAATGATTTGGCTAGCACAGTTGCTAACTATAAAGCTACAAATGACCGAAAAA
    TTGAGGAAATTATTGGTAAAGGAGGAGAACGTATTACATCAAAACAAGTTGATAAACTGATTAAGGAGGG
    TAACAATCAAATCTCTGCAAAAGCATTATCCAAAGTTGGGAATGATTACAATACGAGTAAAGATAGACAG
    AACGTATCTAATAGCTTAGCAAAATTGATTTCTTCAGTCGAAAGCTTTACGTCTTCCTCAAACTTTAGGA
    ATAATTTAGGAGCATATGTTCCTTCATCAATAAATGTCTCGAATAATATTCAATTAGCTAGAGCCGCTTA
    A
  • Molecule Role : Protective antigen
  • Related Vaccine(s): A. pleuropneumoniae DNA vaccine pcDNA-apxIA/pcDNA-apxIIA
9. ApxIIC
  • Gene Name : ApxIIC
  • Sequence Strain (Species/Organism) : Actinobacillus pleuropneumoniae
  • NCBI Protein GI : 52630373
  • Other Database IDs : CDD:154216
  • Taxonomy ID : 715
  • Gene Strand (Orientation) : ?
  • Protein Name : ApxIIC
  • Protein Length : 160
  • Protein Note : biovar: 1
  • Protein Sequence : Show Sequence
    >gi|52630373|gb|AAU84699.1| ApxIIC [Actinobacillus pleuropneumoniae]
    MMLKNDFNVLGQIAWLWANSPMHRNWSVSLLMKNVIPAIENDQYLLLVDDGFPIAYCSWAKLTLESEARY
    VKDTNSLKIDDWNAGDRIWIIDWIAPFGDSSLLYKHMRQRFPYDIGRAIRIYPSKKDTGKIIYLKGGKIT
    KKVAEKTFLQYEQELITALQ
  • Molecule Role : Virmugen
  • Molecule Role Annotation : In the present study, the gene encoding the ApxII-activating protein, apxIIC, was insertionally inactivated on the chromosome of a serovar 7 strain, HS93. Pigs vaccinated with live HS93C-Ampr via the intranasal route were protected against a cross-serovar challenge with a virulent serovar 1 strain of A. pleuropneumoniae (Prideaux et al., 1999).
  • Related Vaccine(s): A. pleuropneumoniae HS93C-Ampr , Actinobacillus pleuropneumoniae apxIIC and apxIVA double deletion mutant vaccine
10. apxIIIB
  • Gene Name : apxIIIB
  • Sequence Strain (Species/Organism) : Actinobacillus pleuropneumoniae serovar 2
  • NCBI Nucleotide GI : 470686
  • Other Database IDs : CDD:30270
    CDD:162558
    CDD:186074
    CDD:73011
  • Taxonomy ID : 34063
  • Gene Strand (Orientation) : ?
  • Protein Name : RTX toxin
  • Protein Note : A sub-family of peptidase C39 which contains Cyclolysin and Hemolysin processing peptidases. Peptidase family C39 mostly contains bacteriocin-processing endopeptidases from bacteria. The cysteine peptidases in family C39 cleave the 'double-glycine'...; cd02417
  • DNA Sequence : Show Sequence
    >gb|L12145.1|ACNNON:3943-6078 Actinobacillus pleuropneumoniae (serotype 2) RTX toxin III (apxIII CABD) genes, complete cds
    ATGGAGTCACAAATGCCTTTTAACGAAAAAATAGATTACGGATTACATGCATTGGTAATTCTCGCGCAAT
    ATCACAATGTTGCCGTAAACCCTGAAGAGGTAAAACATAAATTTGATCTTGATGGCAAAGGATTGGATCT
    TGTTGCTTGGTTATTAGCAGCAAAATCATTAGAATTAAAAGTCAAACGAGTAAAAAAGAGTATTGAGCGT
    TTACCATTTATTCATCTTCCTGCTTTAATCTGGCGAGATGATGGTCAACACGTTATTTTGATGAAAATTG
    ACACCCAAACTAACCGTTACCTTATTTTTGACTTAGAAGAACGAAACCCTAAAGTACTAAGTGCGGCTGA
    ATTTCACGAAATTTTTCAAGGTGGTATGATTCTTATTACTTCACGAGCTTCTATTATGGGGCAATTGGCG
    AAGTTTGATTTCACTTGGTTTATCCCCGCAGTAATTAAATACCGTAAAATTTTTGTAGAAACTATTATTG
    TTTCTATTTTTTTGCAGCTTTTTGCACTAATTACTCCCTTATTTTTCCAAGTTGTGATGGATAAAGTTCT
    TGTCCATCGTGGATTTTCTACACTTAATGTTATCACGGTTGCATTATCTGTAGTGGTTATCTTTGAAATT
    GTATTAAGCGGTCTACGGACTTATATATTTTCCCATAGCACTAGCCGAATTGATGTAGAACTTGGTGCAA
    AATTATTTCGTCACTTGTTAGCGTTACCTATTTCTTATTTCGAAAATAGACGTGTAGGTGACACAGTTGC
    TCGAGTACGAGAATTGGATCAAATACGCAATTTTTTAACAGGTCAGGCACTTACCTCTGTATTAGATCTC
    TTATTCTCTTTTATTTTCTTTGCAGTGATGTGGTATTACAGCCCAAAACTAACTATTGTGATTTTACTTT
    CATTACCTTGTTATATCGCATGGTCAATATTTATTAGCCCAATATTACGTCGTCGTCTAGATGAAAAATT
    TGCTCGTAATGCTGATAATCAATCTTTTTTAGTTGAATCTGTTTCTGCAATAGACACGATCAAGGCTCTT
    GCTGTAACACCTCAAATGACAAATATTTGGGATAAACAGTTAGCAAGTTATGTATCAGCAGATTTTAGAG
    TGACAGTATTGGCAACTATTGGACAGCAAGGTGTACAACTTATCCAAAAAACAGTAATGATAATTAATTT
    ATGGTTAGGTGCACATTTAGTAATTTCAGGGGATCTTAGCATTGGACAATTAATTACTTTTAATATGCTT
    TCAGGACAAGTTATTGCACCTGTAGTTCGTTTAGCACAATTGTGGCAAGACTTTCAACAAGTAGGAATTT
    CTATTACACGATTGGGAGATGTCTTAAATTCACCTACAGAAAATTATCAAGGTAAGCTTTCACTACCAGA
    AATCTTTGGGGATATCGCATTTAAACATATTCGCTTTCGCTATAAGCCCGATGCTCCAATCATTTTAGAT
    GATGTAAATTTATCGGTTAAACAGGGGGAAGTTATTGGGATAGTAGGACGTTCAGGTTCAGGTAAAAGTA
    CTCTCACTAAATTATTACAACGTTTTTATATTCCGGAAAATGGCCAAGTATTGATTGATGGTCACGATCT
    TGCGCTTGCTGATCCTAATTGGTTACGTCGTCAAATTGGTGTTGTTTTACAAGATAATGTGTTATTAAAC
    CGTAGTATTCGCGATAATATCGCACTCACTGATCCAAGCATGTCTATGGAACGTGTTATCTATGCGGCAA
    AATTAGCAGGGGCACATGATTTTATTTCTGAATTACGTGAAGGTTACAATACTATTGTAGGAGAGCTTGG
    TGCAGGCTTATCTGGTGGACAACGTCAACGGATTGCTATTGCACGAGCTTTAGTCAATAACCCTAGGATT
    TTGATTTTTGATGAGGCGACAAGTGCATTAGATTATGAATCTGAACATATCATTATGCAAAATATGCAAA
    AAATCTGCCATGGACGGACAGTAATCATTATTGCCCACCGTCTTTCTACAGTAAAAAATGCGGATCGCAT
    TATTGTTATGGAAAAGGGACATATTGTAGAGCAAGGTAAACATAACCAATTACTGGAAAATGAAAATGGA
    CTCTATTATTACCTCAACCAACTACAATCAAATTAA
  • Molecule Role : Virmugen
  • Molecule Role Annotation : An apxIIIB mutant, in combination with an apxIIID mutation, is highly attenuated in pigs and induces significant protection from challenge with wild type A. pleuropneumoniae. The mutant also was slightly attenuated in mice and induced protection in mice against challenge from the parent strain (Park et al., 2009).
  • Related Vaccine(s): Actinobacillus pleuropneumoniae apxIIIB and apxIIID double mutant vaccine
11. apxIIID
  • Gene Name : apxIIID
  • Sequence Strain (Species/Organism) : Actinobacillus pleuropneumoniae serovar 2
  • NCBI Nucleotide GI : 470687
  • Other Database IDs : CDD:130902
  • Taxonomy ID : 34063
  • Gene Strand (Orientation) : ?
  • Protein Name : RTX toxin
  • Protein Note : type I secretion membrane fusion protein, HlyD family; TIGR01843
  • DNA Sequence : Show Sequence
    >gb|L12145.1|ACNNON:6090-7523 Actinobacillus pleuropneumoniae (serotype 2) RTX toxin III (apxIII CABD) genes, complete cds
    ATGAAGTTATGGATTCTAGGACTTGGGGAATTTTTTCAACGTTATCGTAATATTTGGCGTGAAATATGGA
    AAATCCGCAAACAATTAGATACCCCAGCAAGACAAAAAGATGAAAACGAATTTTTGCCTCGGCATTTAGA
    GTTAATTGAGACACCTATTTCAAAAAAGCCACGGCTGATCGCTTATTTGATAATGCTATTTCTATTTTTA
    GCTATTGTAATTTCCATTATTAGTAAAGTAGAAATTGTTGCTAGTGCTACAGGTAAGTTGGTATTTAGTG
    GACATAGTAAAGAAATAAAGCCTATTGAGAATGCTTTAGTAAAAGACATTTTTGTTAAAGATGGACAATT
    TGTTGAAAAAGGACAATTATTATTAAATCTCACCGCACTTGGCTGCGATGCAGACAAACAAAAAACTAAA
    GTATCGTTAGGATTGGAAAGATTAGATGGTTACCGATATAAGTCATTGTTATATAGCATTGAACACAATA
    GATTACCTTTATTGGATTTTAACCAAGCTGATTTTGATTCTGTTCAGGAAGAAGATAAGACTGGCGCACG
    TCATTTAATTACCGAACAATTTGAGACTTGGCAAAAACAAAAATATCAGAAGGAATTAGCGTATCAACGT
    AAACAAGCTGAAAAACAAACAGTATTAGCAAATATCCGTAAATATGAAAGCGCTAGTCGTATTGAAAAGG
    AGAAATTAAGTGATTTAAAAAAATTATATGATGTAAAGTCTATTTCTAAGCATGAGTTGTTAGCACAAGA
    AAATAGATATGTTGAAGCTAGTAATGAATTGTCTGTTTATCAATCTCATCTCAAAGAAGTAGAAAGTGAC
    TTGCTTAAAGCACAAGAAGATTTAAAGCTTGTTACTCAATTATTTAAGAGTGATATTTTGGAAAAACTAC
    AGCAAAATATACAACGCGAAAAGCAGCTCACTTTAGAACTTGAGAAAAATGAACAACGTCAATTAGCCTC
    TATCATTAGGGCGCCAGTATCAGGCACAGTCCAACAATTAAAAACTCATACTAAAGGTGGCGTAGTAACT
    ACTGCAGAAACCTTAATGGTCATTGCTCCTGAGGATGACGTGTTGGAAGTAAGTGCTTTAATTCAAAACA
    AAGATGTTGGTTTTGTTGAAATTGGACAGGAAGCAGTTATTAAAGTGGAAACTTTTCCCTACACAAGATA
    TGGTTATCTCTATGGAAAAGTAAAAACTATTACTCTTGATGCTATTGAGCACCCTCAGCTTGGTTTAGTT
    TTCAATTCTATTATTGAGATTAACAAGAAAACATTAACAGATGGTGATAAAGAAATTCAATTAGGTTCTG
    GAATGAGCGTTATTGCAGAAATTAAAACAGGAGAACGCAGTGTTATCAGTTTCCTACTCAGTCCATTAGA
    AGAATCTATTACTGAAAGTCTAAGAGAACGTTAA
  • Molecule Role : Virmugen
  • Molecule Role Annotation : An apxIIID mutant, in combination with an apxIIIB mutation, is highly attenuated in pigs and induces significant protection from challenge with wild type A. pleuropneumoniae. The mutant also was slightly attenuated in mice and induced protection in mice against challenge from the parent strain (Park et al., 2009).
  • Related Vaccine(s): Actinobacillus pleuropneumoniae apxIIIB and apxIIID double mutant vaccine
12. ApxIVA
  • Gene Name : ApxIVA
  • Sequence Strain (Species/Organism) : Actinobacillus pleuropneumoniae
  • NCBI Protein GI : 6671152
  • Other Database IDs : CDD:164328
    CDD:116920
  • Taxonomy ID : 715
  • Gene Strand (Orientation) : ?
  • Protein Name : ApxIVA
  • Protein Length : 130
  • Protein Note : Peptidase M10 serralysin C terminal; cl12108
  • Protein Sequence : Show Sequence
    >gi|6671152|gb|AAF23114.1| ApxIVA [Actinobacillus pleuropneumoniae]
    DWGRNSVIDAGAGNDTVNGGYGDDTLIGGKGNDILKGSYGADTYLFSKGHGQDVIYEYSNSASSTSDIDT
    LKFTDIGLSELWFSRENNDLIIKSLLSEDKVTVQNWYSHQDHKIENIRLSNEQMLVSTQV
  • Molecule Role : Virmugen
  • Molecule Role Annotation : A precisely defined DeltaapxIICDeltaapxIVA double-deletion mutant of A. pleuropneumoniae was constructed based on HB04C(-) by transconjugation and counterselection. The results demonstrated that the DeltaapxIICDeltaapxIVA double mutant strain was less virulent than HB04C(-). Despite attenuation of virulence, the DeltaapxIICDeltaapxIVA double mutant remains immunogenic and conferred a similar level of protective immunity to pigs against challenge with a lethal dose of a heterologous fully virulent standard serovar 1 strain of A. pleuropneumoniae (Liu et al., 2007).
  • Related Vaccine(s): Actinobacillus pleuropneumoniae apxIIC and apxIVA double deletion mutant vaccine
13. CP
  • Gene Name : CP
  • Sequence Strain (Species/Organism) : Actinobacillus pleuropneumoniae
  • VO ID : VO_0011021
  • Gene Strand (Orientation) : ?
  • Protein Name : Capsular polysaccharide
  • Molecule Role : Protective antigen
  • Molecule Role Annotation : Following a booster vaccination, a significant (P < 0.05) IgG antibody response to the CP (cps5C), LPS, and HP was detected in the vaccinated pigs. The pigs vaccinated with the CP-HP and LPS-HP conjugates exhibited significantly less mortality (P < 0.05) and significantly greater weight gain (P < 0.001) than unvaccinated pigs. Vaccinated pigs exhibited significantly fewer and less extensive gross pulmonary lesions (P < 0.001) when compared with unvaccinated pigs (Byrd et al., 1992).
14. Lipopolysaccharide
  • Gene Name : Lipopolysaccharide
  • Sequence Strain (Species/Organism) : Actinobacillus pleuropneumoniae serovar 3 str. JL03
  • VO ID : VO_0011023
  • Taxonomy ID : 434271
  • Molecule Role : Protective antigen
  • Molecule Role Annotation : Following a booster vaccination, a significant (P < 0.05) IgG antibody response to the CP, LPS (wzxE), and HP was detected in the vaccinated pigs. The pigs vaccinated with the CP-HP and LPS-HP conjugates exhibited significantly less mortality (P < 0.05) and significantly greater weight gain (P < 0.001) than unvaccinated pigs. Vaccinated pigs exhibited significantly fewer and less extensive gross pulmonary lesions (P < 0.001) when compared with unvaccinated pigs (Byrd et al., 1992).
15. sd
  • Gene Name : sd
16. ureC
  • Gene Name : ureC
  • Sequence Strain (Species/Organism) : Actinobacillus pleuropneumoniae serovar 2
  • NCBI Protein GI : 303253439
  • Other Database IDs : CDD:183894
    CDD:30031
  • Taxonomy ID : 754254
  • Gene Strand (Orientation) : ?
  • Protein Name : urease subunit alpha
  • Protein Length : 572
  • Protein Note : urease subunit alpha; Reviewed; PRK13207
  • Protein Sequence : Show Sequence
    >gi|303253439|ref|ZP_07339581.1| urease subunit alpha [Actinobacillus pleuropneumoniae serovar 2 str. 4226]
    MALIIPRSQYVATYGPTVGDKVRLGDTDLWATIEQDFLTKGDECKFGGGKSVRDGMAQSSTATRDNPNVL
    DFALTNVMIIDAKLGIIKADIGIRDGRIVGIGQAGNPDTMDNVTPNMIIGASTEVHNGAHLIATAGGIDT
    HIHWICPQQAQHAIENGITTMIGGGSGPADGTHATTCTPGKFNIERMFQACEALPVNIGFFGKGNCSMLE
    PLKEQVVAGALGLKIHEDWGATPAVIDAALKVADEMDVQVAIHTDTLNESGFLEDTMKAINGRVIHTFHT
    EGAGGGHAPDIIKAAMYPNVLPASTNPTRPFTVNTIDEHLDMLMVCHHLDKRVPEDVAFADSRIRPETIA
    AEDILHDMGVFSIMSSDSQAMGRVGEVVTRTWQTADKMKAQRGALGDEGNDNFRIKRYIAKYTINPAIAH
    GICQYVGSLEVGKLADIVLWKPQFFGVKPEFVMKKGFISFAKMGDPNASIPTPQPVFYRPMFGANAKANT
    ESAVYFVSQASVDANIKAQYGIQKETLAVKGCRDVGKKDLVHNNATPEITVDPERYEVRVDGEHITCEPA
    TKVPLAQRYFLF
  • Molecule Role : Virmugen
  • Molecule Role Annotation : Deletions were introduced into the ureC and apxIIA genes of an Actinobacillus pleuropneumoniae serotype 2 strain by homologous recombination and counterselection. The double-mutant contains no foreign DNA, is highly attenuated, protects pigs from homologous challenge upon a single aerosol application (Tonpitak et al., 2002).
  • Related Vaccine(s): Actinobacillus pleuropneumoniae ureC and apxIIA double mutant deletion vaccine
1. A2M
  • Gene Name : A2M
  • Sequence Strain (Species/Organism) : Homo sapiens
  • NCBI Gene ID : 2
  • NCBI Protein GI : 66932947
  • Genbank Accession : ABBA01012257
  • Protein Accession : NP_000005
  • Taxonomy ID : 9606
  • Chromosome No : 12
  • Gene Starting Position : 9067707
  • Gene Ending Position : 9115961
  • Gene Strand (Orientation) : -
  • Protein Name : transcript variant X1
  • Protein pI : 6.39
  • Protein Weight : 149502.37
  • Protein Length : 1474
  • Protein Note : Also known as A2MD; CPAMD5; FWP007; S863-7
  • DNA Sequence : Show Sequence
    >gi|568815586:9067707-9115961 Homo sapiens chromosome 12, GRCh38 Primary Assembly
    AGACATTGACCAGAAAAAGTGTTTATTCATCAAGTCTTTAAAGATACAAAAACACGTGTCTTCTGTGGAG
    CTCTGAGAACAGGACTCCAGCAAAGCACTTTTCAGCCTTGTGGTCTTCAAGCATTTCCAAGATCTGTGAC
    ATTGGAAAGAAAAAAAATTAAGACAGATTTGGGTCTCCATGAGCAGAGAGTATTCTTTCCCTTAGTTCTG
    GGTAGCATAGTGCCCAAGGAAACCTAATCAGTACAGTGGGAAACCAAATCTATTCCAAATCATTACCCAT
    AAGCAATCCTATGGACTCTCTGAGGTTTGACAGAGTCAGCGGCCCTCTCCAATAAATGTGTTTTTCTATA
    ATAATGTGCAGTGTGAGTAATTTGATGTTTTTGACAAATATTTACCCAGCGTTTTATGAAGGAGAAGGTT
    TGGGGGGCAAGCTGAAGGAGCTCTGACTGCCTTTTGGAGGAGTGTGAGTGGCTTACCTTTGCTGCAAGGA
    GCATTGTACTCAGCAATTGCAAACTCATCTGAAAAAAAAAAAACCAACAAAAAACCAGAAATCATTACAT
    GAAGTGAGAAAGAAAGCAAACATCTGTGGGATACAGGCCAAGGAAGGAGTTTGTTGTGGGAAGAACAGTA
    TTGTACCAGAAACATAAGCATCATTCATCTGATGTGTTGCTTGCTACCTTATATTCTCCTTTCAAATATA
    GTGTTTTATATATTCTTTTATCCCAAACTCCTGAAGGACTCTTCAGATGTTCTACTTACTTAAATCTCAT
    ATTGTCATACAGTCCATTTATCTGATGAAAAGGGAACTAAATATCCCACAGGTGTTTTGTATTAGTAGTT
    GATAAATTTATAATGTAGGTTGTTTCTATAATGTATGTATAAGAGACAAAAAGAGGGGCATCAGACTTGA
    TGGAGACAAAATGAAGTGATAATGTAATTACTTAATGTAATAAATAACCCCAACACATCTCCTAAACCTG
    AATTTCTGTGATCAGGAATTAAATATTTCTACGTGAAAATCACTCTCACTCACCCGTCTCGTAGTAATCA
    TAGACTTTCACTATGGCTGGTTTCAGATCTCTTACTGGGACATCTTGCAGAACCGTGAAGAACAAGCTCA
    GTGTCTGATTTGACACCTGGAAAGCAAAAGTCAATTAAATGACCTTTCAGGAAGCTTTCTTCTCTCTTTG
    AATTAGTTTAAGTATTCACCTGTTTTTTGGGGGAAAAGCTTTATTATCCTACAGCACACTATAGGGTCCT
    TAATTAAATTGTGCAGAAATCTCTCAGAGGGGATGATAAATTCTCCAGATTATTTCTTAAATAACTAACA
    TAACGCATATACCTCTGTCAATGCCAATACATAGCTCTGTTCATGGGGGTAAATGATATATACTTTTTAT
    TGGATGACTAACTGTGTTCATTTTTTAATATCCAGAGATTTGGTATAGTTTGACTTCTCTGTCTTGCAGG
    AATACTTGCGTAGACTCAAAGAGGCATCTAAAAGGTTGATCTTCCCAGGCTTAAGTTTCATAAAATTTGT
    AAACTATAATTCCCATCAACTTATGCATGAACAAACTCTCCTACCACACATATCTATAAAATCAGATTGG
    TTTACTGGTGAACTGGTCTTATTATTTTATGGTTCAAGTCCTTGTGGAGACTGACAGTTTTTTTTCCCTT
    TCATATTGTTGTTGAGTATGTTGATAATTAACTAATAATTGACTAATCACAAGGGCTACTTCCTTATATT
    ACTGGGTATCCAAAAAACTTGTCAACATAATAAACACATTTTGTGAAGATTTCTTGCTTATGGGATTTTA
    GGAATATTCATATCCCAAGATCATAATAAGCCATTTCTACCTGTAATCCCTGGTGAAATATAAATAAAGG
    AGAGAAGAACATTCTTTTAGAGTTAACATTAGAATTCTCAAATTTCTAGCATGGAAGTGATGTTTCTAAA
    ATGCATTTACCTTCCCAGATGTTCCCTTAGAGGATTATTATCTACGTTTTTTTGCAAATAGACTGGAAGT
    TCTCTTACCTTATCAAGGTAAATCAAGACATGGTTGCTGCTGACTTCTGTCCGGCTCACATGGTTAGATC
    TTTCAAGCTGAAGAAAATTGAAATACCACAAATGTTAATAAATAGATGAAACCCCTGGGGGATCCAGAGA
    AAAAATCTTTGTATTGCCAAAATAACCCTGAGGGTAGAATTCTTCAAATGCTTTTTGTAAGGAAATATAT
    AATGTAATACAATTAAAAATGAAAGGCATAAAGGGTTTACTATTAGTATAACACACGACTAATGTGTAAA
    GTTATTTATCTGAAGAAAAGACAGCTAAGTATCCTAAACTTGGTAGTTACTCATATTTCAAAGTTCAATT
    CAGTTTCTATTTCTTCATAAGGAGTTTTTTTCCTGTTTTGCTTGCTCTATTTTTCTTTTAACTTCTGTAA
    TACTTATAGTTTATGCCAATCAGTCATTTTCATATTTTTCACTGGTTCGTGTTCATATCTTTTATCTTCT
    TAGCAAAATTCTAAGCTAAGAAGAAATTGAAAAGCAGGAACCTGCTGACAATCTTTTTCTTGTAGTCATG
    GGTCCTAGCACAGTGCTCTGCACATGGTCAGCATTCATACAAACACATGTGATTATATTTCTGAGCTGGA
    AAAAGGATAAGGCTTTGATAGAGATTGACTTTAATAGTATTTTGATATTAGTATTGTCTTATGCTGGGGA
    TACATACATCATTAAATAAAATAGGGCAGTCTGGGGTTATGATAAACCTACCATTTTCACTGTTGGCTTC
    AGGGGAATGAAGCCAGAGACCATCTTCACATCAACGATCGCCATGTTGGAGGCAGAGCGGCTCCCTGTGT
    AACTGAGGATCCAGGGGAGGAAAGGATTAGGGTTTTCTGTGTTTTTAAATATTTTCTTCTTCTATGTCCA
    TGTTAAGCATTCCACAGCTCTAAAAGGTACACGTAAGTCTTCCCAAATATCTTATCCCAGTGGTTTGTAA
    AAGTGGTGTGCGTAGAATTTTTAGGAAGCCTGTTAAAGCCACAGAGACCAGGCCCTACTCTAGACCTGAT
    GAATCAGAATGTCTAGTGTATGGGGGCTGAGGATCTGCATTTTTTTTTTTTGATGGAGTTTTTCACTCTT
    CTTGCCCAGGCTGGAGTGCAATGGCATGATTTCGGCTCACCGCAACCTCCGCCTCCTGGGTTCAAGTGAT
    TCTCCTGCCTCAGCCTCCCGAGTAGCTGGGATTACAGGCATGCACCACCACGTCCGACTAATTTTGTATT
    TTTAGTACAGACGGGGTTTCTCCATGTTGGTCAGGCTGGTCTTGAACTCCCGACCTAAGGTAATCTGCCC
    GCCTTGGCCTCCCAAAGTGCTGAGATTACAGGCGTGAGCCACCACGCCCGGCTGAGGATCTGCATTTTAA
    GAAAGCTCTACAAGTGATGCACAGGCTGTTTGTGACCTCTAATCTATGCCAGTTATTTTATTTATTTGTT
    TACTTTTATTTTCTTTCAGCTGACAAAATGATGTATATTATGAATAATAAATTTCAGTCTACATTATTTA
    CAAAAACAACAATAATTTATAATGTTTACTTAAACACTGGCTTTCAACAAAACACTTAGAAACACCTAGC
    GTCTTGTCCCCGAGAGTTTGCATCCGAAATTTGGACTGCTTAATTTATTTTTTGGCATATATATACACCA
    AATTAAATGAAAAAAATATATATATATATACTTAACTTTTAGGTTCAGGCGTACACATGCAGGTTTGTTA
    TATAGGTAAACTCGTGTCATGGGGGTTTGGTGTACAGATTACTTTATCACTTGGGTACTAAATTTTGCAC
    TGAATTGTTGTTTTTTTCTGATCTTCTCCCTCCTCCCATCCTTAACCCTCAAGTAGGCTCTGGTGTCTGT
    TGTTCTCCTTTTTGTGTCCATATGTTCTCATCATTTAGCTCCCACTTACAAATGAGAACATGCAGTGTTT
    GGTTTTCCATTCCTGTGCTAGTTTGCCAAGGATAATGACCTCCATCTCCATCCATGTTCCTGCAAAGGTC
    ATGATCTCATTCTTTTTGATGTGTGCCAGTTATTTGTTACATGTATTTTGTCCTAACTTTTTTCCTGAAT
    AGTATCTCCAACCAAGTAATAATATGTTTGTGGAGAGAGAGTTTTTTATCAGATCTGCAGCTATTTAAAT
    ATTGATGCATATAAGTGTATCTTCCCCCCAAATTAATTATATCATATATAACCTGAGATTTATTATTGCC
    AGTAGGAATTCTAACTAAGGAGTCACTACTTCCTTCTATTCAAACTTTTCAATAATTCTTCATCTCTATC
    TTCAATAAAGCAAGAATAATATAGGATTTCATTACATAACAAGGAATTCAGGTCAGAGGCAATGGGTAAT
    ATTTACTTAGTTGTTATTGAGGGAAATTATTACCCTTTAAACAATGAGGTTGCACAATAGTAGATCCTGG
    ATGGACAATGTAAACCCCATCATTGAGAAATTATATCAACTTAAGAGAATACATTGCATTTTTTGTGAAT
    AGTGCAAATATCTACTGTTGCACTGCTTTACTTAAAGAGGAGTTCCCGAAAGAAGACTGGTGGTTATTCT
    TAGGATTAGGTGATAGAGTCAGAAGGTCTTACCTGACACTTAGGGAGATTTGGAAGCTGGTGTGGGCTTT
    GGGTTCATCACAAGTTTGAGGCAGAGTCTGCACTCCTAAAGCAAAGGGGAACTCTTCCTTTTCTGGGAGA
    ATATTGTATTTCAAGGATGTCTATAGAACATCAAAGACAAAATCAGTTTTTTGCCCTTTCCCAGAATTCC
    TGTCCACGTCCCTAACCCTTTCTCTGATCTGTCCCATTGTTTTCTGAGTTAGGACTTCTCAGAGAGAACA
    GCTGCTGTCCTCATCTGTCCTGTCCTCACCTGCCCAAGAGTCTCACCTGGAGGTAGACACATCCTTCTCC
    TGTCACTTTCATGCTGTATTCCCCAGGCAGCTCTGGCAATGAGACCTGCTGCAGTAACAGGCGGTTGTTG
    TTGTCCACTTGGAATTTGCTGGAAAATGTCCCTGAAGACTGGATAGTCACCTGTGCAGCCTTCCCAGTCC
    TGGTAAATGTGGCTGCTCCATATTTGGACAGAGCATGGAGAGCCACCACTGTGTCCTGTTAGAGACAGAT
    GAGTGAGAGAACCCATTGGGCATTATAAGGCTTTGAGGGCTGAGATATTTTTCAAAGACCCATGTGAGGG
    ACTTTGATTTCCTACTTCCCTCACTACTTAAATATAGGAGCTGTAATTTCAAGATTGATTATAATCTTTA
    GATAGTTGAAGAACCATAACATTCACATATCTACCTTGGACTGTATGCCCTACATGTTTATTTTATTTAT
    CAGAAAATTGATCCTTTTCTCTCTGTAAGTATATGTGGGACATGGAATGTGGAAAACATGTGAATGGCAA
    TCAGTTATTGGCATCAGCTATTTGCTAACATGTCTTTCTTGACAACTATGCTCATATCTGCCACCACTCA
    AGCAAAGTTCAGTCTGGCTCTCTCCCATATTAAACTTTGCTCAGTTGCTGAGGTCAGTGTCCTGGAGAGA
    ACCCAAGGTTCTTTGCAGCCATGAGCAGAGTCATGCTTCAGTTTCTCCAAAAGAAATAAAAGATGAGTAT
    TATAATTATCTTCTGGATTTAATGACATTTTTATTTGGCTTTCCCTTTTTATTGGATGATTTCCTGTTCC
    CAGTGAATGTCTGCTGGTATGTATCCTCAACAACATTAAACCTGCTTGGCCTCCTTCAAATAATATGTTT
    CTAATAAATGAATGATAACCTTTAACAAAATTACTTAGTTTTTTTCATTCATATGATAAAAGATTAATAC
    AATGGAAATAGGATGGAATGCCATTTTGTCACATTTTTTCCGTTTTAGGGTATATAAGCAAACATCCGAA
    TTCTCTGAAGATAAAACTTTCTGCTGTGTACATATCAATGAATTGTTCAGATACATGGTTATATAGCTAG
    ATTTAGGAGGTTGGATGAATAAAGACAGTTCTTTATGCTCCTGATGAATAAATGTTTGTGTATTAAGAGG
    TGAATAGGGGGCTGGACGCAGTGGCTCACACCTATAATCCTAGCACTTTGTGAGGCCAAGGCGGATGGAT
    TGCCTGAGCTCAGGACTTCGAGATCAGCCTGGGCAACATGGCAAAACCTCGTCTCTACTAAAAATACAAA
    AATTAGCCAGGCATGATTGCACATGACTGTAATCCTAGATACTTGGGAGGCTGAGGCACTAGAATCGCTT
    GAACCTGGGAGATGGAGGTTGCAGTGAGCCGAGATTGCACCACTGCATTCCAGCCTAGGTGACAGAGCAA
    GACTCTGTCTTAAAAAAAAAAAAAAAAAAAAGGTGAATAGGGGAAGCACAAAATGTGTAGTTTTGAGAAA
    AGCCTATTAAAGGGGACCTAAGAATGGAGAGATTTGCGAGGAGAGGTTGTTGGATTTTGAGTAGAGTTCC
    TTTGAACAGTTTGAAAGAGACCCTAGAGCTTTCAGGAGAATGGCAGGAGACATCTCCTTCATTCTGCTGG
    TGCTCCACTGCTGCTTGAGGAAGGCAGTGTTGTTATGGGAAATAGGAGATGGATAATAACACAGCGATGG
    GGTGCCGGGATTCTGAGCATTTCCTTTTGGCCCTCTTTGACTTTGATTTGCTTTTATTTCCATTATCCTT
    GGATACTGAAAACTTTTCCTCATTTCAAGTTACTGTCATCTGTATTTTTTTCTTCTTGGATGTGTTTGTT
    TCTTTTTCATTCAAATCTTTTGCTACCTGAAGGTGAAATAAAAGGTTTTGGCAAATCACCAACCTGGGTG
    GAGGAGAAACCGCCCTGGGCATTCTGCTGCTTCGTGATCCACTTCACGATGTTGGTTGCAGAGGTCAGGT
    CCTCCGAGGTTGGGGCTGGCTGGGCCGTGAGATAAGCGAGGAGCACATAGGATGTCATCTCCACCTCAGC
    AGAGGGAGCCTGGGGTTCGTAAAAATGCCCCACTGGTGCCTTGGGTTTCTGAGGGCGCTCCCAATGGACA
    GAGTTGTCTTAAAGATGAGAAAAAGATATTTATAAGTGCCTACATATTTATATTTAATTCAAGGTGAAAG
    TACCCAAGCCAGTGCTGAAATGAGAATTGCATGCCCATTATAATCCCCTGTACTGTATGTAGATGCTTTT
    CCCTTCATGTAAATCTGAATTTGGTGTGTTTTAGCCCAGCTAAGGCAGATATTGATGCTGGAGTCAGAAG
    CCTATGCCTATCTATGTTTTCAAATCGTTGCTATAGTCGGGTTGGTGGGAGAAAGAAATGGCACAGCAGT
    TGTCTCATTGAGTCTGGCTATTTCCTATTTATGATGTTATATTAGACTATAGCTGGAGTTTGTTACATTT
    CCTATAAAGTGTGTCTTTATCAGTTTCTTGTGAGGATCACAGATCACATGAAATAGTAGAGTTTCTACTC
    ATTAAATTGAAAAAAAAAATACAGTAATCCTTGAAATACCAAAGGCTGACATGGATGTGAAACAATGGGA
    ACCAATATACACTGCTGTGGGAAGCCAACTGCTCTTTGGCAATATTCAGCAATGGGGAAGATGTGCTTAG
    CCCACTAATCAGTCATTATGCTTCTCAAAGCATAATTTGGAGAAACACAGATGCACAAGAAAACAGGCAC
    ATGAGTATTCATTGCAGCATTTTTTTTAATAGCAAAACATAAAAAAAATCCATGAGGAGAAACTCGATGT
    GTAAATTATGACTAATTTATAGACTGTAATATATTATGCAGCAGTGAAAATTAATGAGCTGACTCTGCAC
    ATACTTGTATAGGGGTCTTCATCAACATAGTGTTGAACTTAGAAAATAAAAAGCAGACAAACAAAAAACT
    AAGCTGCTGAAATATTCAAAAATGTTTCTCTTGTTTCTTGAATAAGAGAGAAATAAAAATCTTGTTTCTC
    TCAAGATGAAACACATGTGAAGATTCTAAATATTAGTTAGGGATACATATGCAGCAAAAATATAAGGAAA
    TCCATGGAATTGTAACGACAAATTCAGGATAATGATTGCCTCTGGAGGTGGTAGTGATACACAAGGGATT
    TATTTTTCCCATGTCGGTAATTTTCTATTTCTTAATCTATGAGAGAGTTGTACAGTGTTTTCTAGTTATA
    GCTTATGCCCTTTTGTATATCTATTATAAAATATTTCTCAATACATTTTAAGAAGAATTAATGGCTGTCA
    AAGGAAAAAGGTAGACTGTTTTAATGGACATCTTGTGATTTGCATTTTATTTTCTATTTCTTCAACCGCT
    CATATAGACCATGGTATATGTAGAAGTACTGTGTGAATATGAGTAGATTGGTCAACATTATTGGTCTTAT
    GAAATGGGCTTAAAATACCAGAATTCATTCAAGGAGTTACTTCCATGATTATTTTCTTATGTTTAAGGTT
    ATATAATGTACCAGGTAAGGGTTGGGCATATATTGAGAACTGATATGTTAATTGTAATAACTAGTACTTT
    TATAGGATTTGCTATGTGACATGTACTCTACTAAGTGTTTTACGTGTACCAATTCATTTATTACAACAGC
    TCTGTGAGGTAAATACTATTATTATCCCCATTTTACTGATGAAGGAACTGAAACACAAGGAAGTTAATGA
    CTTTCCCAAGCTCACATAGGTAGCACATTTTGGAGCTAGTCTCCAGAGAATGCTCTTCTTACCAGCATAC
    TATAGAGATGGTCCCTGACTTACAATGGTCCCACTTAAGATTTTTTAGACTTTACAATGAGTTTGTCAGG
    ATATAACCCCATCGTAATTGAGAAGCATCTGCACGGCCTCTTGTCTTCTGATTTCTGATTTGATTAAGAA
    ATCCTAGATTTTATAAGATGCAATATGGACAAAATATATATGATATATAGAAAAGAAGAGAGGAGACAGG
    GATCACAGGAGGTAGGAGTGAGGGATTTTTGCCATGCAGTTCTTGATACAGAGGATGGGCCAGGAGAAGG
    ATCTCAGGTGTGCTCTCACCTTTCTTCACAGCTTCCTCATTAAGTGACTTGAGTACTTCCTTCCTCTTGT
    CCTGGTTACCTGCCAGGGCAAAAGCATAGGCCAGCAGTGCTTTGGTATATACATGGCTGCCATGGTCCCC
    TTCTTGTGCTGTCTTCCAGGCTGACTCCAGGCAAAACAGGGCATTGCGGACAACAGGGTGCTGTGAAGGC
    AGAACAAGAAGGGAACTAAGCTATGTAAACAGGCATATTAGCATTCCCAGGCAAATACACGGATCACAGC
    ACAGAAGTTTTTCAAACGCATTTTTCCAACGATTCCTCATTCTTATCAATAGATATAATATTATTTTTAT
    CTGTTATTCTCACATTCAAAAGATACATCTTTAAGATTCTTTTTTGAGTGGTGGTGGTAAATAGCCTAGG
    GGAATTACTGGGAAAATTTAAACCTTTCTTAATGGGCTGACCAAATTCTCTGTAGATCTGGGAAGCACTG
    GCATTGCATAGAAAGAAAGCTGATGCTTTGCTTTTAATAGGATAGTATTTCAGACAGCAGGTCAGCAGTT
    TCATTGCATCCAGAACTCTCCTTCAGCAGAGGAATGGGGTGGTACCTACAGTGACTGTGAGAGGAATCTC
    CAGAAGGGCGATGGTGATATAGGCGGAGAGGGTCACTTCATCTTCTACTCCTCCCTGTGAATACGAGAGA
    GAGATCTGAATAATTCAACTTCTGAGAATGCTATTGATTAGTTCTTTCTATTCTGCTGTGTCATGGAATT
    CATCCTTACCCATATCCATCCCTATAGGGCAAAGTATCACAATCAACTTTTGTTCTATCCCCTCTTTTTT
    GGTGCCATCCAGGTTTTATTTTCCTCTGAGGCTTTCCCTTAGAATTTTTCACATTATCACTTCCTATCCT
    CATCCTTGCAGATATCATAATGGACAAATCAAAACTAGCTCCAGAATGATTCACCTTTATGGCATTGTTG
    AGCAGTGACCCAGAGCTCCTGAAACAGCCATTGTCCTTCTGCCTCTGGGAGAGCCATATGAGGGCTTGGG
    TAATGTGTGCTTCATCGATGAAGATGTAGGCTCGAGCTTGGGCAAAAGTCTTCAGAACAAAGGCTGTGAG
    CCTGACCAGGGAGGAAGCAATCATGATGTTTATGTTGTCAAAATGGTTTTATAACCACTTCACAGCAACA
    GGCTTCATATGATGTGAGTTAGCTAACAAAATTCTTGTGTACTTAGAGAGCTTATCTCCTTGATTAATCT
    TTAGTCTGCCTGATTAATCTGCTAGATGTAGTATTTCCTTGCTAACTCTCTCTTTGCCACAGGGAGTCTT
    AGTATATTTAATTTTATTTTAGTTCCGAGATACATGTGCAGAACGTGCAGGTTTGTTACATAAGTATACG
    TGTGCCATGGTGGTTTCTTGCACCTATTGACCCATCCGCTTAGTTCCCTCCCGTCACCCCACACCTCCCA
    ACAGGCCCTGGTGTGTGTTGTTCCTGTCCCTGTGTCCGTGTGTTCTCATTGCTTAACTCCCACTTATGAG
    TGAGAAATGTGGTGTTTGGTTTTCTGTTCCTGTGTTAGTTTGCTGAGGCTGATGGCTTCCAGCTTCATCC
    ATGTCCCTGCAAAGGACATGATTTCATTTCTTTTTATTGCTGCATAGTATTCCATGATGTATATGTACCA
    CATTTTCTTTATCCAGTCTGTCATTGATGAGCATTTGGGTTAGTTCCATGACTTTGTTATTGTAAATAGT
    GCCGCGATAAACATACATGTGCATGTGTCATTATAGTAGAATGATTTATATTCCTTTGGGTATATACCCA
    GTAATGGGATTACTAGGTCAAATGGTATTTCTGGTTCTAGATCCTTGAAGAATCGCCATACTGTCTTCCA
    CAATGGTTGAACAAATTTACATTCAAGTAATTTGAATGTACTTATTTTTGTTAGTTACTTGAGTAAGGCT
    TTAAATCTCTCTTATCTTTAGAGATTTCCTTCTGCAGATGACAGAATCTTCTCACATGCAGCTCTTACAT
    GCAGTAAATTTTAATATTGCTTTACTTTAGGGATTTAAAAAAATGTTTTATATTACTGTTTCCTTCTCAA
    TTGTTTCCTCTTATTAATGAATTCATTCCAAGTCAAGCTGAATGACTTAGAATGTGACAATATAAGCTGA
    AGTTAACATGGTTTATCTGGAAGTTATATATGTTCACTGGCAATATCATTTTAAAACTCATGGTTATGGT
    AGCTACTTTAGAAAGAATAGCCATTTCAGAAAGAAAACCTTGGTACATTATTAATAGGATCCATTTTGGA
    AACAAGCCTAACATAACATGTATTTGGATATCAGGTTTTCTTCTGATTACCTTGCTAATCTTGGTGGATT
    TATCTAGTATAACAAACCATCGGTCTGATAATACATATCTGATAGTGTTGCTGTGAATATAATTGAGGTA
    ATACATGTAAAAGAGCTGGCACACAAAAAGAAGCTCAAAAAATTGTTCTTTCCTTACCAGGTGTTGCCCT
    GGTTCCTGCCATATCGCTCCCCAAAGGTGCTGTAGGAGCCATCATAGTGTTTGTAGTTCAACTGTCTCTG
    GTAACCTGGAAAGGAAGATTAACGAAACAGCACAATGGATTAATGTGCATGCTGAGGGTGGAGAAATTAC
    TAAAAGTACCTTGGCTTCTCTTGTGACATTTCTTAAATTTTGTTGTCATAGATTAGGAGTTTCTGAGCCT
    TAAATATTTTATTGGAGGTTGGAGAGTGGATAGTTTCCTTGAAATTAACTATCATAGCAGCTATCATAGT
    GAGCTAAGCTAATGTATCATAATATTCATAAGTAACTGAAACCTACTGGGAAATCCAGTTGAAATAACAT
    TCAAGTTTTCCCTTACTCAAGTAATCACTCACCAGTGTTGAGATAGCCAATGGCCTTGGACTTGATCTCT
    GGAGTAAGCTGCTGTGTTTCATTTAGATAATCCAGTACATAGATGTTAGGAGCAAAGAGGACCATATTCT
    GCTCTCCACAGCCATAGGGCATCTGGAGAAGATTTTGTGTGTTTTGCATGGCAGAGCCTAATATGTCTCC
    TAGAGAATGGGAGAGATGGGAAGTCATAAAGCTTGGAGATTATCATCTATCAAAGTCATTAAGCAGAAAT
    AATTAGTTGAGCTTAGAAATTGAGAATTTTTAGGAAGGATGATTCTTCCAGGGATAGAAGTATGATTGAA
    AGCAATAAACAAGCCCAAAGAAGAAGAGAAGAAAGAAGTTAAAATTATAGTATTATTTTTAGTAAATATT
    TATGGGAAATAAAAATAGTATAATAGAAGCTGTTAATGCCCGGATCCACTAGGGGCTGGAGACTCACCCA
    AAACTGAGACAGAAGCTCGGGCAGATTCTTCTACCACATTTGGTGGCAGTTTCAGGGATAATTCTTCAGA
    AACCTCACCACCTAGAGAAATAAGCAAATCAGACATATGAAAATTATTTCTGCATTATATCTTTCTAAAC
    AGCTCTATGACCCAGAAGCTAGGACTATGCCTTATACCTAAACTCCTCCTGAAAAGTTGTCCGCCTTTAA
    TACAACAGTAATATATAAATATCTTAAAAATGGGCTACTGGTTTCCTGTGATTTTCAGAATTTCTTCTTC
    AATGATCTTTTCAGTAATTTATGTAGAATCGCATGCCAAATAAGACAACAAATTGTAAGAGCCTGGTGAT
    GTTGTGATTCAGTTGCTGTCATCTGATGGCTGACACAAGACGGTTATCACAAGATGATCTATTTAGTTCC
    CACCACCAACTTCCTTACTCAACAGTTGTATGGAGAAAACACCAACAACAAAGCCCTATAAGTATACATA
    AGTAAAATTACCCACAAAAGGTCCTGAATTCTATAATAATGCTATGTTCTTTCTTCAAATTTCATATACA
    AAGATAGTATTTTTATGGAAAACCTAGAAAATTGAGGGTAAGGAAGTGAAGACGAACAGTATTTGATGCA
    TTGAAAATAAAATAATTACTACTCATGAACATACATTTACTTCCATTTTAATCTTGGTAAATTATATGAA
    ATGAAAATGAGAGAAGTATGGTATTTATTTTCATACTGAATCCCTTCCTCATAAAATTCATATTAAAATG
    TTTCTTAAAAAAAAGCCATAAGCATAGTTAAAGGACAAATGGCTAAGCAAAATAAAATGCTTGACTCACT
    GAAAATAAAATAACTGCTACTGATGAACATACATTTACTACAATTTTAGTATTGGTGAATTATATACAAA
    TGAAAAATGTAGCAGTTATCTTCCCTTTCTCATAAAATTAATGTTAAAACTTTTGTAAAAAAACAGATGC
    CATAAACAAAGTTAAAAAAATGACCAAGTGAAAGAAAATATCTGCAACATATTCAACAGATAAAGAATTA
    GTGTCATACAACTGAGTCAGAAAAAAACTACTCAAAAGACAAATAGCCAAAGATTATAAAGGCAATTAAT
    AAACATGCAAAAATAAGGTCATCCTTGTAATTAAGAAAATATAAATTAAAACAACAGCAAAATTTTTTTT
    TCTAAATTTTTGGCAAAAATTAAAATTAAAAACTTTGATGGTAACAGATGATGGCAAATATAACACTATG
    GAAATGTATCCTGGTGGAAGTATAAATGCATTTAGGTAATAAAGAGGAGGATTTTCGTTTTGTAGTTCAA
    TTAAAATTCAGAATAAAGCAAGATGATGGACTAGAAAGTCTCAGGATCTTCCTCCCTCCACAGAAAGTCC
    AACTAGCTGCTATCCACAGATCAGAAAATCTATTTTGAAAACCCCAATACTTGGAAACAAGTCTAGGACA
    CCTACATTGTTCATAGAACTGAGTGAAGACAAATTAGAAGAGTAGGAAGAATGGTCTCACTTTGACCATG
    TCATTGTTCCCACTCCTTTAGGTTAGCACAATGCCAGATAGAGAGGAGTTCCCTGGGGTCACAGTTTCTA
    CAGCGGGGAAGAGAACCAGGGGTAGACATCAAGATTCTCAAGTATTCAAAGACACTTCTCAGGAAGCCCA
    CTCTGGTCTCATTTCACCGGGAACACAGGAAGTAATGAGGTCAGGTAGAAACAAAGAAGGACAGAGCTCA
    CAGTGACCAGCATGTGGATTTTGGTAGTGCTTCTGTGTTTCTGCCAGCTATGGCACCCATTCAGAGATAC
    CAGCCAACTGCATAGCCCATCTGCAAAACTGAGCTGGTCACTTTCAGAAGCATGTTGGGAAGTTCAACCT
    AGCTTGAGTTTCCAGATGGCCAGTCTGTAAGCCCAGCTTCAGAGGTTATGCCAGTGACCCCACCCAGGTA
    GCAGACACCACCTCTGTGTATTTCATAGAAGTGTAGGGGTTAAATATGCTTGACCTGGGAGTCTAAAGGG
    TAGCTCAACTCAGCCAAAACCTCACTCCATGGCTCCATTCACAAGGGTGGGGTAGCAATCCTCTATTATG
    CACATCTAAGAAGTATAGTATTTATTTCTGCCCATCCCAAGCAACAACTCTACCTAACCTCACAGCCCGG
    ATTGTGACCCTGCCTAACTTCAGGTCCCAAATAGCAGAATCATCCAGCCAGGCAACACATCTTGTGACTG
    GCCTGACCAGAGGCCATTGTAGTACCCAGCCAGCAGCTCCACCAGATAGCAGAGACAAAACAGTGATCTT
    GCCAGAGAGCAAGGCCCAACCAGTGGCCCTACCAAACATAAGAGCAAAGGCAGTGGCCCAGCCAACTAGG
    GAACCCATGAAAACTCTGCCTGCTCAAGGTTGTCACCAACTGATCTTTCCAGAATTATAGGCTAGACTAA
    ATAGCAAACCTTTATCCCTGCCAAAGAATACCTATAAAAACCAGAAGAAGTGGCTTTCTCCTCAAATGCA
    CAAATAATGCAAGGACACAGGATTATGAAGAATCAAGGAATGATGACACCTCCAAAAGAAACTGGCCAAG
    CTCAAAAAATTGACCTCATAGAAATAAAGATCTATGAAATCACTGACAAAGAATTCAGAATAATCCTCTT
    GAAGAAATTCAGTGGACTACAAGAATATATAAATACAAAATTAAATGAAATTTGCAAAACAATACACCAA
    CAAACTGAGAAGTTTGACATAGAAACGATTGTGAATAGTGCCACAATAAACATATGTGTGCATGTGTCTT
    TATAGCAGCATGATTTATAATCCTTTGGGTATATACCCAGTAATGGGATGGCTGGGTCAAATGTATTTCT
    AGTTCTAGATCCCTGAGGAATCGCCACATTGACTTCCACAGTGGTTGAACTAGTTTATAGTCCCACCAAC
    AGTGTAAAAGTGTTCCTATTTCTCCACATCCTCTCCAGCACCTGTTGTTTCCTGGGACATGGATGAAGCT
    GGAAACCATCATTCTCAGCAAACTATTGCAAGGACAGAAAAACAAACACCGCATGTTCTCACTCATAGGT
    GGGAATTGAACAATGAGAACACTTGGACACAGGAAGGGGAACATCACACACCGGGGCCTGTTGTGGGGTG
    AGGGGATGGGGGAGGGATAGCATTAGGAGATATAGCTAATGTAAATGACGAGTTAATGGGTGCAGCACAC
    CAACATGGCACATGTATACATATGTAATAAACCTGCACATTGTGCACATGTACCCTAGAACTTAAAGTAT
    AAAAAAAAAAGAAATAGAAACAATTATCTGCAACTTAGCTAATATTGTAAAATAAAAAATAAATAGAAAC
    AATCAAAAAGAAACCAAATAGAAATCCTGGAAATAAAGAATACAATGACTAAAATTAAAAATTCAAAGTA
    AGCTCAACAGCAGACTTGATCAAGCAGAAGAATCAGTGAGCTCAAAGATAGAGTATTTGGAATTATCCAG
    GCAGAAGAGCAGAAAGACAAAAGAATGAAGAAAGCCTATGGAAATTATGGGACATCATCATGAAATCAAA
    CCTTTGCATTAAAGAAAGTCCAGAAGAAGAAGAAAGAGACTATAAAGCACATTTAAAGCAATAATGAAAA
    AAAAGAGTATAGAATCAGAAAAAAAAAAAAGCAGTAAGGGCTGAAAACTTCCCTAATTAGGGGATAGATG
    CTAGCATCCAAGTACAGGAGGCACAGAGGTCTCCAGTGAAATTCAAATGAAAGAACTGCTCACTAAGACA
    CAATAATTAAACTATCAAAAATCAAAGACAAAAAAAAATTCTGAAAAGCAGAAGGTGATAAAGAAACATA
    TCACATACAAAGGAATTCCAATACAACTATCCATGGATTTTTAGCAGTAACCCGGCAGGCTGAGGGAGAG
    TGGAATAATATATTCAAAGTACTGAAGGAAAACAAAAATAAAAACAACATGGAATCAAGAATTCTTCATC
    CAGCAAAGCTGTCTTTCAGAAATGAGGGAGAAATAAAACATTCCCAGACAAACTAAAAGCTAAGGGAGTC
    GTCAACATTAGGCCTGCCTTCAGGAATTACTGAAGGGAGTTCTTTAAGCTGAAACAAAAGGCCCCTAATT
    AATAATACAAAAACACAAAAGCACAAAATTTAATGGTATAAGTAATATAGAGCCATATTCAAAATACTCT
    ATGACTGTAATGATGGTATTTAAAGCAATTTTATCTCTAGTACAAGGGTTAAAAGACAAAATATTAATAA
    CAAGCATAGCTAAAAATAAATTGTCAATAAATACACATTACAAAATATGTAAATTCTGACATCAACGACA
    TAAAATGTGTGTGGGGGAGTGAAAGAATGCAAAGTTAAGTTGTTATCAGCTTGAAATTGACTGTAAACCC
    CATAGTAACAGCAAGGCAAAAATCTTTAATAGAAGCACAGAACAAAAATTTTAAAAGATTCAAATCATAC
    CACTACAGAACATCAGACCACAAAGGAAGACAGCAAAAGAGGAACAAAGAAGCAAAACAACCAGAAAACA
    ACAAAATGGTAGTACCAAGTCATTTTTAAATTAATAATTACCTTGAATGCAAATGGACTAAATTCCCCAT
    TTAAAAGACATAAAGTGACTGAATGGATTTAAAAAAACAGGACCGAAACATATGCTAATTACAAGAGACT
    CACCTAATTTTTAAGGACACACATAGACTGAAAGTGAAGAGAAGGAAAAAGATGTTCTAGGTAAATGGGG
    ACTAAAAGAGAGCAGGGGTAGCTATACTTATATTCAACAAAACAGACTTAAAGTCAAAAACTGTACAAAG
    AGACAAAGGAGAACATTATATAATGATAAAAGGGTCAATTCATCAAAAGGATATAACAATTGTAAATCTA
    TACATATCCAACATTGGAGCACCTAAATATATAAAACAAATATTAAAGGTTGAAGGGAGAGATATGTTGC
    AATGCAGTAATATCAGGGAATTTCAACATCTCACTTTTAAAAATGGATAGATTATCCAGAAAGAAAATTA
    ATAAGAAAACGTTGAACTTGACCAACACTTTAGATCAAATAGACCTAATGGATACATACAGAACATTTCA
    TCTCAAGCACACAAGAAACATTCTCCAAGATAGATCATATTTTAGCCCACAAAACAAGTCTTAGCAAATT
    TAAGAAGACTGAAATCGTTTCATCGTATCAAGTATCTTTTCTGACCACAATACTATGAAAACTAGAAATG
    AGTAACAGGAGGAATTAGAAAAAATTCACAAATATATGAAAATTAAATGACATGCTCCTGAACAGCCAAT
    GGGTCAATAAAGAAATTTAAAAGGAAATTCAAAAATATCTTAAGACAAATGAACATGGAAATGGAGCATA
    ACAAAACATAGCATAAAGTAAAAGCAATTCTAAGAGGAAAGCTTATAACAATAAACACCAAAAAAGAAGA
    AAAATTTCCAATAAAGATACTAATGATATTCCTCAGAGAAGTAGGAAAAGAAGAACAAAGTAAGCTCAAA
    ATTGGCAGAAGAAAGGAAAATAATAAAGATTAGAGCAGAAATAAATCAAATAGAGACTAGAAAAAGAATA
    CAAAAGATCAATGAAACTGAATTGTTTTTTTAAAAAGACAGACAAAAACTTTTAGCTAGGTTAGCTAAGA
    AAAAAGAGGGAGGCCTTAAATATATAAAATCAGAAATGAAAGAGAAATTATGGCTGATACCACAGAAATA
    CAAAGGATCATAAGAGACTATTATGCATAAGTATACACCAATAAATTGGATGATCTAGAAGAAATGAATG
    CATTCCTAGACATATACATACTACCAAGACTGAATCATGAAGAAATAGAAAATCTGAACTGACAAATAAT
    GAGTAAAGAGATTGAATCAGTAATAAAAAGTTTTCCATTAAAGAAAAGCCTAATAACTGTTGGCTTCACT
    GCTAGGTTCTACTTATCATTTAAAAAGAATGAATACCAATCCTTCTCAAACTCTGCCAAAAGCTGGAAGA
    GGAAGGAATACTTTCAAGCTCATTTTATGAGGCAAGCATTACCCTGATACCAATGTCATAAAAAGACATT
    ACAAAAAAGAATTACAGACCAATATTCTTGATGAATATAGATGCAAAAATCCTCAATAAAATACTAGCAT
    AGCAGCTAAGCATGGTGGCATACACCTGTGGGTGGCTGAGGTGGTAGGATTGCTTGAACTCAGGAGTTCA
    AGTCCAGCCTGGGGAACACAGCAAAAGCTCATCACTTAAAAAACTAAACCAACCAACCAACCACACGAAA
    TAAACTAGCAGACTAAATCCAACAATACAGTAAAAGGATCATCTACCTAGTCAAGTGGGATTTATCCCTG
    AGATGCAAAAATTGTTCAACATATGCATATCAATAAATGTGATACATTACATTTACAGAATGAAGGGTAA
    AAACCATACCATCATCTTGTTAGATGGAGAAAAAGTATTTGACAGAATTCAATGCTCTTTTATGATAAAA
    CTCTCACCAAATTAGATATACAAGCAATGTTCCTCAACACAATAAAGGGCATGTATGATAAACCTATAGC
    TTACATATTTAACAGTGGAAAATTGAAAGACTTTCCTTTAAGATCTGGAACAAGACAAGGATACCCACTG
    CCACCATTTATTTTCAAAGTAGCACTGGAAGTAGCAGCAAGAGCAATCAAACATGAAAAAGAAATGAAAA
    ATATCCAAATTGACAAGGAAGAAGTAAAATTTTCACTGTTGGTGATGACATTATCTTATATATTGGAAGC
    TCAAAAGACTTCACCAAAGGGCTGCAAGAACTTATAAATGAATGCAATAAAGTTGCAAGATACAAAATTA
    ACACATAAAAATCAGTAGTATTTTCTGTTCACTAACAATGAATGATTGAGAAAGAAATAAAAACAATCCC
    ATTTACAATAGCTATAAACAATATGTAGAAATAAATCTAATCAAGGAGGTGAAATGTACAGTGAAAATTA
    TAAAACACTGATGAAAGAAATCAAAGAGAACACAAATAAATGAAAAAAAATACAATGTTCATGGATTGGA
    AGAATTAATACTGTTAAAATGTCCACACTACCCAAAGCAATCTACATATTGAATGTAATCCCTAACAAAT
    TACTGATGTCTAGTTATGGAATCATAAGTGTCATCAACAGATGAACAGATAAAGAAAATTTGGTATATAC
    ACATAGAATTCCATTCCATTTTAAAAAACAAGAACATTCTGCTATTTATGGCAGCATGATTAAATCTGGA
    GGAGATTATGTTATGTGAAATAAGCCAGGTATAGGAAAACAAATATTGCATGTTCTCATTTGTATGTGGA
    AGCTAAAACATTCAAATTCACAGGAGCAGAGAGTAGAATGGTGGTTATGGAGGATGAAGCTGGAGTAAAA
    GGAAGGTGGTGTTCAAGGGACACAAAGTCTCAGACAGAAGGAATACATTTTTTTCTTTTTGGGATCTATT
    GCACAGCATGGTGAATATAGTTAATAATAGTGTATTTTATGTTTCAAACTTGCTTAGAGAATTTCAAATG
    TTCCCACCACAAAACATAAGTATTTCAGGTTAGCTTGATTTAATTATTTCACATTTTATTCATAAATCAT
    AACATCACTTTGTATGCTGTATATATATACAATTATAAATGGTCAGTTTACAATGTAATTAAAGAACAAC
    AACAACAACCAACCAAACAAAACTATGCATATCTATGCATATGCTCTCTGACTCAGGAATTTAACATTTT
    GGATCTACCCTAGCTGTACTTGCATATGTTCATTAAGAGGCTCTTATTATAATTAACTAACTAATTTTTT
    AAAGCTAGTGAATAGTTGAAAAAACCAATATAGGAAGACTAAAACACTAAGCAGAATTGTGATGTATTTA
    TATTATAGGCTTAAAAGCAATTCCAAGGAAAGACCTAGATCTACAATTTAACATAGGTAAATCTCCAAGA
    CAGGATGCTGAGTGAAAAACAAACAGTGATTTATAGAATGAAACTTATACAGTTTATTATATGTAAATCA
    CACATCCACAAACATATACATCTACACACATTATTTCTAAAGATAAATGGATGGCAAAGTACTGAAAAAT
    GTTCAGGGGGGTGACATTAACCTTTAATGTGATGTTCTGTTTTTTTTTTTAAAGAAGAATATATTTTTGT
    AAAATTAGGTAGGCACATTTGATTATGTTGGTAATTAAAATGAAATTTAGTAAAACTATTTGATGATTAG
    CCCTTACACTTAAGAGATACATCAAAATGAGTAACTCAGAAACGGAGAAGTCATGATAAAAGGATTGAAT
    GATAGGTAATCAGTTCACTTAAATGTAGAAATAAGGCTAAATGACAAGGAAGTTATGTTTTTAGGGTACA
    TTTTATGTGTTACATCTTGGCAACAGATGAATAGATAAAGTATTTAGTACGAAAAACATACTAAATTTGT
    AGGTAGTCAGAGAAAGGCAATGAACCTATGTATGATGTTGATTCAATGTTGGTTTGGGGTGAGAATGCAT
    TGTGCTGATAAATTTCTTAGTGTATTAAATTTTTCCTGTAAGTTTCTTCAGTTCCTTCTCAAATGTTTGA
    TAGGGCATTAGGTACGGGATAGGCATCAAACAGATATTAATATGACAATAATGGTAAAGGCCATGACAAC
    TTTTCCTTTGGGAAATAGCAATCCCGTTATCAAATACTGTTGGCTATGTAAAACTCAGTGTTAGAATGCC
    CTAAATTCAGCCGTCCATAGTTTGCATTATTAATTATTTAAGGAAACATCCCTTTTGGTAATATTAATTT
    TGATTCTACCCACTTTCTACCTATGCCCATACCATACACTGGTCAGAAAGTCATTTGGTGTTTGCACAGG
    TGCTGCAAAAGATCAAGACTCAGAATGCAGGCCTAGGGATATTCTGATCAAATTACTTGGCCTTCCTAAG
    AACTATTTTCCACATTTGTCAAATGCATTGATGGTGCTTCAGGTCTTAGATCACAGAGAAAGTGTAGGAA
    TAATATATAAATGTTCTTTGAACTTTAAATGTTAGACTTTAATGTTTTTTAAATTATGATGGTTGACTCT
    TACCTGATGGACAAAGTAGGGAGTTGAATGTTGTTTCCTTCTCTAGTCCTTCAGGCTTTAGGTAAAAGAA
    AGAAAAGCTAGTGAGAATGGACTGACCTTCAGAACATGTGTTTTAATGGAGGGACCACAGATATTTGGAT
    AGTGAAGAGAAGGATTTGAACTGTATTATCTAAGAAAATTCTGTACATATAAGCAATATACGTAGGAGGT
    AGTTCGATGCATGAACAAATGAAGGAAAAATTAAGAAAGTAAATCAAAGGGAGGCTGTGCATGGTGGCTC
    ACACCTATAATCCCAGCACTTTGGGAAGCTGAGGTGGGCAGATCACTTGAGGCCAGGAGTTCGAGACCAG
    CCTGGCCAACGTGGTGAAATCCTGTCTCTACTAAAAATACAAAAACTAGCTGGGCGTGGTGGTGCACACC
    TGTAATCCCAGCTACTCAGGAGGCTGAGGCACGAGAATTGCTTGAACCCAGGAGGCGGAGGTTGCAGTGA
    GCAGAGATCCTGCCACTGCACTCCAGCCTGGATGACAGAGCAAGACTCTGCCTCAAAAACAAAAACAAAA
    ACCCCCAACAAAACAGAAAGTAAATCAAGAGAGAGATAGGACAGAGAGATACATGAGAATAATATTATAA
    AATATCCATATATTATTATATTACCCACATATATTATATATTATTGTGGACTATATATTATTTAGGTTTA
    CTTACTTCAACCAACAGAGGCTTGATGACTGTGTCTTTCCTTCCGTGTTCAGGAACTGAAGGCACCTCAG
    TCCCACACAGCTCTTGAGACTCTAGTGCCTCTGCGCTCACAGTGAAATTCACATTTCCTGAAAAAAAAGG
    CCAGTAGAAATGAATAGCATCTTCCCCTTCCTCCATGCCTCCAAACTCAAGATTTAGAAATGTTCAATGC
    TCTGTAAACTTTTGTTTTATTAAAACACAGAAGCAGGAACGGAAACATGCAAATGAGAGGTGAATGATAC
    TGAGAATTCAAGGTGGAGATGTTGAGGATATCCTTGTAGGCATCTTAGAATAAAAGGCAAAGGAAAAAAA
    TCGCAGCATCTAGTGGTCTTTTCCTGAAGGAAGTAGCACTCAATATAAGGTTCCCACTGCCATATACAAT
    CTTTGAGTAGAGAATTATCTCTAGCAGTTTTTTGCTCACCTAATGACTTTGGGGTTACTGCCCAGGACAC
    AGTTTGCCGCCCGTTTGCACAGATGCAGTGAGGCGCTTGTTCCTTCTCCACTGGGACAGCTAGGAAGGCG
    GGAGAGGCTTCCAGCTGCACACTGACCTGAAACCACACATAAGAAAGGGAGTAGAGAGGGAAGGAGAACA
    GAGGGAGAATGGGTTTGAAGTAAAGCATCTTGAGGAATTGGTTAGATTTCAGGTTGCCTCACATTAAAGA
    TGATATAAATGAAGATCAAGTACCTGAAATATATTTAAGTGGATCTTAATGTATCAGATTTACACAGGTA
    AATCTGAGTTATTTTGGAAGCAACCTGCTCTTTCACATGATGGGACCAGAGGAAGGGAAAGGTAATGATG
    GCATGCTGCTCTACCTCTTTGGTCTGGAACTTTGACCATTCCCCCGGATCTCCCGGCAGAGGATACAAGA
    CTTATAAACAGTTATGGTTTCAGCAAAGGTAAGATGCTCACAAACGACTTTTAAAATTAGGTTTAATATA
    ATGAATGAGCTACGAAAAGTTAAATATATGAGATCGTAGAGGTTGAAACCTTGTAGAATCTGTCATGAGA
    ATAGAGTACTGACTTTTGTCACCCCTGTCAGAATTCTTCATACCACACAAATACTAACTTTGAAGGGTTA
    TAAGGGCTAGAAACTTGCTTTATGGTGCAATTTGGTATTTGGAGATCTCAAAACCTGTAACTTTTACAAT
    CATGAATATCTAGTAAGCATATTTTGCATACAAGAGTTTGCAGTATTCCTAGGAATGCAACTTTTAATTT
    ACTTGATGGCTACCTTGTATTTAATTTAGGAAAGAGATCCTTACCCGGATGCATTTGGGAAGGTAGTTTA
    GGACCGTGGCCTTGAGTGTGAAGGCCTCTCCACGAATCACAGAGTAAGGCATTGTGAGCTCCACAAAGAA
    GGGCTGGAAGGCTCGGAGAGAGGCAGTGGAAGAGATACCAAGTCCAGCATCTTCAGACAGGCAGAAGGCC
    CCTGCCTTCCACTCGGTGATGGTGTCAGGGACTGTTACTCCTACCTCAGCCACACCTGCTGAGCTGGAGA
    GGAGTGTAAGTGAAGAACAGAAAGTAAGCAGTTAAATACATCCTTTCTAGGGAGAGAATCCCTAGGAATG
    ATTCTACTGCCATGACTTAAAAACACTCAATAGAAATACCAATAATGGAGAGTATAATCAAGGATTGAGA
    TGGGAATATAAGTCCTTTTCTTCCAACTGTTTTTAATTGTAGTATATCAGAATTGTAGCATAATCTTTTA
    CAGACAATATAGTTATAAATTTAACATGCAGTTTGAGATTGTTCTTAAGTTCTCTTCTCTAAAATGCTAT
    TTCACTGAGGGCTGCTTTATAATTTTAAATTCTACTACTATTGTGTATAGTGGGCCTGACTGAATCTGTA
    TTTAACTCAGGATTGCTAACCAGTGAGGGATCTCTTACAGGCAATCTGTGCAGTCCTTGGTTGTATTAGG
    TCCTTATGCCCATTTTCTGGAAATCTTAGAATTAAATTTGCACAAAATTTTTTAGCTCTGATGCAATTAG
    ATTAGTAAGACTTCTTAGCTGATGTATTTATGTTAAACATCTCCTGAAATTAGCAATGCGTTTGAAAGAA
    GTACTTCAGTACTGAAAAATTAGCTAACAGCTCCCAGTTTCTCCCTGGAAGGGCTTCGTCTACCCACTTT
    ATCAGTTGCTGACTGAGTGTTGGGCTTCTAACTAGCCTGCACTGGGAAGCAGAAGGAGCAGGGAAACGAC
    AGACTTACAGGAAGGCTGAAAAGCAGTAAGGGCACTTCCCATGCCTCCTTCCCCTGGCTCGCCCCACGGA
    TAAAACCAGGTCTGAACATACCCCTGGGAATAATTTGTCTATGCATCAAGCACCCCAATGTTTACAGCTC
    GTACCTAAGGGTCTGGCTCCTAAATCACCTAGCTCCAGGAGTAGATGGAGCTCTGCAATCAGGAGTCTCC
    CTAGACCACAGTGAACACAGAGGTGGTTTTAAATAGGCCTTTGAGCAATGCCAGTGGCTACCCGCCCCCA
    CCACCCCTTGGCTCAATACAGAGAGAACAGGCAAAATCTCCCATTTCTGGCTGTCTCCTAAGGGTTGGGC
    TTCTAAACAGCCTGCATCTGGGAGTGAAAGGGAGAAGTAAACAAGACTCCAGGAGCCTGAAAGGGAGTAT
    ATGGCCATTCCGGCTCTTTCTCTCCACCTGCCCCACTGGCTTGTTTCACGGATAAAACTCTGTTTCCAGT
    GTCTCTCTGGAAGGAATTTGTTCACTCCCCTTGGTGCAAATGTACATTGCCACAGCCATTATGGAAAACA
    GTATGGAGATACCTCAAAAATTTTAAAATGGAAACTACCATATGTCCCAGAAATCCCTCTTCTGGGTATA
    CTCCCAAAGGAAATGAAATAAGCACCTTGTAGAGATATATGTGTTTTGGTGTTCATTACAGCATTATTCA
    CAGTTGCCAAGATATAAAAACAACCTAAGTGTCTGGGTGAATGGATAAATAAATTGTGGCTATGCGTAAT
    GGAATATTATTCAGCCTTAAGTGGAAGAAGATCCTGCCACTTGTGACAACATGGATGAACCTGGGGGAGA
    TTATGCTAAGTGAAATAAGCCAGACACAGGACAAGTACTGCGTGATCTTACTTGTATGTGGATGCTAAAA
    GAAGTTGAATACATAGGAACAGAGAGTAGAAAGGTGGTTGGGGTTGCAGGGATGGGAAGAAGCAGGTCAA
    AGGGTACAAAGTTGAAGTTACGTAGGATGAATAAATTTAGAAATCTAAAGTACAGCATGAGGACTGCAGT
    TGATAATACTGTATTGGATACTGAAACTTGGCTAAGAGGATAGATTTTAGGTGTGCTTGCTATGCACACA
    AAAATAGGTAACAATGTGAGCTAATGGATGTGTTAATTTGCTTGACTGTAGTAATCATTTCACTATGTGT
    ATGTGTATATATATATATCAAAACATCATGTTGTACATCATAAACATATAAAACAATAAAAACAGAAAAA
    CTAGCAAAGAGTTGAAAATAGTCAGGGACCTCTATTGTTGCATATTGCATATGCAGGAAGTTACTTACTT
    TACCACCACCAAATCCCAGATCCATGTCTCAGGGAAGTACTTTCGTACGGTCTCCGTGTGAGGCTCTTCA
    ACATGCACCAGGCGTGCATGGCCTCTTCCCATTACATCTGACTCTATGGTGAGTGAGGAAGAAGACATTA
    CAATAAACATACAGATAAAGCTTATGAGAGAATGTAATAGTTGCCACCAAAAAAAAAAAACAAAAAACAA
    ATCGTCTGATGAAATAGAGAGGAAGGAGGAGAGGGCGCTTGGGTCATCAAGGAAAGCTTCTTTGAGGAGG
    AAATATGTTGGTTGACTTAACAAGCCAGGAGAGGCCGGGCGCGGTGGCTCATGCCTGTAATCCCAGCACT
    TTGAGAGGCCGAGGCCGAGACGGCGCCACTGCACTCCAGCCTGGGCGACCTGAGCGACAGAGTGAGACTT
    CGTCTCAAACAAACAAACAAACAAACAAACAACAAAAAAAAACAAGAATAAGCTAGGAGTGAGCTGGGGG
    AGGAGGGTCATAAGCAGAGGCTATAGAGACTTTGGCAGACAGAGGCTTGGATTGTCTGTGGGGCACAAGG
    ACTCTGTGGAGTCCTATGTGGCTGCTGCCGCTGTGCTAAGTGGAGAGTGTTATGACACAAGGCCGTGAAG
    AGGGGCAGGAATCCAAGTTTCCATTGGTGTGCTCCTCTGCAGGAGCAAGGAAAAACTGACACACATGTTA
    TGACAGTGGCTTTGAGAAACAGTTTACAGGAGGGAAAAGTTTAATAATAATAAAGACTTTTTATTCTCTA
    TCGTGTCAGAAGATTACAGTTAGTTTCCTTCACTCTATTCACCTTCTATTAAGGGTGTGCTGGTCAATAT
    TTCACAACCAGCTCTCTGGAAAAAAAAATTGTGCATATATATATATATATATATATATATATATATATAC
    CTATACATGCATAAATTTACTATATATCTTACTGATAAAGGATTATACAATATAATTTATAAATAATAAA
    ATATGCGGTAGGCTTCATAGTGAATTCCATAGAGCCAACTGATTCTCCTTAAATGCTTTCCTTGATTTTT
    GACCCTCTTTTCTCTGTACTCAACTTATCCTCAATCATGATTTGACAATATGGGACTATAAATAAATGCT
    TGATTACTATTCAGTTCATCAAAGAAGTAGCTCATGTCATTGATGAATGTGTGTGGTTCCAACATAAATA
    TTGGCTGATGTTTTGGGGAAGTTGAACAGCTTTTTTACCTGACTGATGGAGAATGTAGTGTTGTTGTGTG
    AACATTTCTATTTCTGTTTACACTAATCCTACGCTTGGAAACTTTCTCTTTCCTTTCAGCCTAACACATG
    TACCTCCATTTACTAGAATTTGAGTTTATATTACTATATGACCTTGAGCAATAACCTTAATATGTATGAC
    TCACATGTCCTTTTTGTTTGTTAATTTTTGTCACATTGTATCTTTTAAAAAATTTAAAATTTGGTGAATA
    TATTAGGCAATATATATTTATTAATTTTTTGTTTACCATAAAAACCTACACGTAGACCTTCAGGTCCATG
    CATTTCATACTGTTGAAGCTGTGGACACATTTTGGGTTTACGAATCTTTGAGTTGGTGAATGCCTTTAAG
    CCCATGTCCTGCAAAGAAAATTATCATCTAATCAGTAAATATATATTATAAAATATACATAGGTAGCTAC
    TTCTTTTTATTGAATTTGACAAGCAATCCAGTTAACTTAAATTAAACTTTTAACATTCACATCTGTAGAA
    GGGATATTTATATACTTAAGGATGTAATTTTATTCTTTTCATTAAACTCTTTCAACAGTAATTATTTTGT
    ATATGTGATTTTAATAAACATCACAGAGCAATTAATTCCTGTGAAAAAAGAAAACTTGAATAATACTCCA
    AAAAGTTCCAGCTTAGTTCAGCAGCTTTTCGCAGATCTGCTGCTATTAGTAGAGAAGCCACTTACCTCTT
    TATATCCCATTACCCTCAACTAGGACATGAAGGCATTCAAATACAGACTCTTTTCCATGTGTAATATTTT
    AAGAAGCTTAAGATCAGACCATCTGCAACATAAGGAGTTTACCTCTAGGAAGCTGTACATATCCTTTTCA
    TTTGTACTTGATACTGGAGTATATGTGATTCCATTAATATAGACATTATGACGATTGATGCAGTCTTCAT
    TGTCCTGGTCATTCAAAGGCCCAGGGAAGCCAGTGAGGTCCTTTTCTGGTAGCAGGTTGTAAACCTGTAC
    AAATACGAAAGACAAAAAGGCAAACTTATTTGTGACTTTTTTTTTTTTTTTTTTTTTTTTTTTTGAGACG
    GAGTCTCGCTCTGTCGCCCAGGCCGGACCGCGGACTGCAGTGGCGCAATCTCGGCTCACTGCAAGCTCCG
    CTTCCCGGGTTCACGCCATTCTCCTGCCTCAGCCTCCCGAGTAGCTGGGACTACAGGCGCCCGCCACCGC
    GCCCGGCTAATTTTTTGTATTTTTAGTAGAGACGGGGTTTCACCTTGTTAGCCAGGATGGTCTCGATCTC
    CTGACCTCATGATCCACCCGCCTCGGCCTCCCAAAGTGCTGGGATTACAGGCGTGAGCCACCGCGCCCGG
    CCTTTGTGACATTTTTTAAGTTCAAGGAGAAAAGAACAGGAATAATGAACATCCGTCTTTCTTTGAGCGC
    TCTCATTGCTTTGATGATTTATAGAGGGTATGGATCATTTTCTTGACACAGTGTTTTCATTTTGAAGATG
    TTTAACAATTTTAGGAAAAACCAAACACTCCATCTAAGTACTCATCTAAGTACTAAGTACTACTTCGTTA
    CACAGTTCAGAGGCCACCACTAGACTCAACACTGGAATGAAATAAAACTTTATTAATTGAAAAATCGAGG
    TCTGCTGTATTTGTGACATTGCACAAAGCTTTGAGATGATAGAAGGAAATGATAGTTTTGTGCTTATCTT
    GTGGTGAGCAACTGACTCAGTATATAGCAAAGTTAATGGGTGCATGTTCTAAACAGGCACACGAATAGGA
    ATATCCTCTCTCCACAGGACACGATGGCATTTTCTTAGGTATTATTAGTGGAAACAAAATGCTCCAGATC
    TTCATATAATACATACAACTTTAACCGTAAAATTCTAGGCATGATGCTTTCAAGTCCATTTGAAGGTGAG
    AAGTAAGACAGACATAAAAATTGTGATAGTTACATTGCATTATATTCGTAGGAGTTTAAAACTGATTGAC
    AAGAACAGATGAGCAAAGTGAGTGGTGTGATGAAATTAGACACTTTCTCAAAGTATCTTAACATTTCTCA
    TTTCCTCTCCCTGCTTTTATTGTGGCATATTGCATAATTATTGTCTGTCTCATCTATGGGACAGACAATG
    TAAGTTCCATGAGACATACATTTTTATCTTTCCAGTAGCTCACTGCTTGATCTCTACTAGCACTTAGTAT
    AATACCTAGAACACAGTGGGCATTAAATAAATGTGTTGAATGAATGACCTACACTAGTAGCAGCAAAGTG
    TTTCATTGGTAAGACACTAATCAAAGCTTGAGAAGAGTTGGTGTTTTAACTCAGGTCTACTACTAAATCC
    CTGAAAGCTTCTGCTTTATTATTAATAAATAGGACTGTCTGCCTTATCTACCCATCAGGGTTGTTATAAT
    TTGAAAAATTCTTTGAGAATTATAAAATTATATTATAAAAGTATAAAACATGTATATTATTTTCCCACAT
    ATCCTCTTGAGATCCTACTGTTGAAATGTACTTGATGCTTCCTTTAGAAGTATTCTAATATTTTCAGTTA
    AAATCCCATTAGAAGGACAAACAAGTAAGAAACATTTTTTTATAAGTTTCTATTTCAGGAAAATAAAAAT
    ATAAATTTCTCTTTGTTAAATTATGGTAGTAGAAAAAGCTTCAGTAAGAGGTTATTACAATTGGTCTCAT
    ATTAAAAATTTGAAGATGTGAGCAAACTACTATTAGTACATGCTATTAACTAGTTTTTGTTGTTCCAATT
    CAGCATATCCTTTTGGTATTTCTTCTATTAGCTCCATATTTTCAAAGAGTTCAGTTATCAATGTCAATTA
    TATAGTAGTTCAATTTCCCTAAGTTTTGTTCCTCAGAGATAATACATATGTTCCCTCAGAAATTATGTGT
    AGCTTGGGACAGCAATTATTTCAACTGATGTAATTCTTTTTTTTTTTTTTTTTTTGACACCAAGTCTTGC
    TCTATTGCCCAGGCTGGAGTGCAGTGGCACGATCTCGGCTCACTCAACCTCTGCCTCCTGGGTTCAAGTG
    ATTCTCCTGCCTCAGCCTCCTGAGTAGCTGGGATGACAGGCGCCCGCCACCACGTCTGACTAATTTTTGT
    ATTTTTAGTAGATACGGGGTTTCATTATATTGGCCAGACTGGTCTCAAACTCCTGATCTCAAGTTATCTG
    CCTGCCTTGGCCTCCCAGTGTTGGGATTACAGGCGTGAGCCACCAGGCCCAGCCATAATTCTTATAAAAA
    CGGAAATCTGATATTTCATTTAGCCTGTATAGCCTCAGTGCAGGTATATGTATCACTTTTGTTAGGCTTT
    TTGAAATGTTGAAACTAGTTTGACAACTCCTATTACTGGTTGCATGGAGCTCTGGGATTTTAAGGACTTT
    GGGTACTGTATGGTCTTTTTACATTTTAAATATATAATTTTCGAACGGCTTCCCCGGTTGCTCAGTTAAT
    CCTTTCTTTTATTTTGCATAACTGCACTTCTCATATAGCACTTTTCACAATCTGCTCTGTATTCTCTCCA
    GCCTTGCATATACGGATGTCCTGACACAATCAGAAGATTTGAATTTGTATTGCTATGTGCTCTTGAGCAA
    GAAGCTTAACGTGTCTGAATAACGAGTCCTTTTTTGGTTGTTAATTTTCATCACATTTTGTCCTTTTAAA
    AATTCTTTTATTTGATTCAATATATTGTATTTTGTTTATTTTTTTAACTGCAGAAGTGAGTAGTTATATA
    TATATATATATATAATTCCTTACCAATGAAAGCCCACAAGAGAGCTCAAAGCAATTAATTCCTAGATAAA
    TATAATTTTAATTGATCTTATCCTACTTATCCAGTCATTTTTCCTTGCTCTGAGCCCCTGGCACGGCCCT
    TCTTGATTCCTGAGGCTGCCAGGAACTCACCGAGGACGCCGAGAGCTCAGCATCAGGCTTCATGAGCAGC
    ACGCTTTGGTCCACAGCACGGAGGGCGCAGACGGACTGAGGAGCCGCTGTGACTCGCAGGTGGGCGTGTG
    AGGCTGGGAGACTTTGTGATGGGCTGAAGCTCAAATCCACCTGTGAAATTGGAACAAAAGGTCAGAAAAG
    CAAATTTCCAGGTTTACCCATGCATTCACTCGCATTTGCAGAGTGTTCCTCATGTACAATGTATAACCAC
    TCTGCTTGACTTCGTGGAGGGTTGGCATTGTGTCAATCCTGAAGCTTTTTGTTCCCTACACTTCTCTTTC
    ATTCTCAAGAATCACTTGAGCCAGTAGAATTATTTCTAATTATTCTTATAAATTACTAGTCATGAAATGC
    CATTTTTATCTTTGATACACGGCAAGGTGCTCATTAATTGTGGTGGGACATGCATGGGTCTAGGTGCTCT
    TGCATTTTTCTTTTGGCTCCTTAGTAGTCTTAAAAACTTGCTTACATATTTCCCAACGTTTCATCATAGA
    AGAAGGGAATAACATACTCTGAACTGTGTAATTGGTAAGAGGCACTGCTAGGAGAAGTTTGAGAGTTTTG
    TATAAGGAATCAAGTGATTCTGCTTCTTAGTGTGACTCTGCCACTACCTTGCTGAATGTCTCTGGGGAAT
    TTGTTTAACCCTTTTGGCCCTAATGTTCACATGTGTAAAACAAATGATAACAATAGTAACTTCTAGTTTT
    ACATGTTGAAGATTTTATGATCTAAAACACACACCTTGTTGGCCAGACAATTTTCAACATCATATTTTGC
    AGAATCCCCAATCACGTCCCCGGTAGGTAAAACAGCATAGATGAGCAACCGAGCGACAGGAGCAATGTCT
    GACTTCACAGGGATTGAGATGGAAAAATGGCCCTTCACTGGGGCACAAAGAGAATGAGAGGAAGCCATCA
    TAGGTTAATGACTATTTCCATTAGTAGATGTAACAGACATAATAAACAGTAGATGTAACAAATGGATGAT
    TACCTCTAAGGACTCTAGCTTTAGAAAAAAACCCTATCAAAGCCCTCTCTCCTCTCATAAATCATATACT
    CTGTTGATTTTTCACCAGCTTTAAGCTTCAGCTCACCCACCTAATGTTCTGATGCTAGTTAAGGGTATGA
    AGACATCCAATTCACTACCATTTGATCTAGGCCTCATACTAGAAGAGGGAAATAGAACCTGAAGTTATGA
    GATGTAAGAGTAAGAGTTCAACACATTGTATTCATGCTACATGCTCTTTAACCTTCTATGAGCCAATATT
    TGGTTCTTTCTCCATAAGAGGAACACCATGCTCCTTCAGAAGTTATTTTTTGATGCCTATGTAGATTCTT
    TAGAGGTTGCATTAAGTTTCCTTCAGTCTGCTATGGTGCTTCTGTGTTGCTACAAGAACTAATGAGAGAG
    TAGATTACTGATACTTTTCTAGCAAGTTATCTTTTAGAGTCTACTAGACTAGGAGCAACCTTCAAAGGTT
    TATATTATGACATATTTGCTCATGGCAAGAGTCCGCTTCAGTCGGTAGAAGTACCCAATTTAATGCTTCA
    TAAATCATAACTATCTTGAAAGGGAAAAAAGATATTTATAATAATATAAATGACATAAATGACTCAGCAC
    AAGACCTGGTATATACAACATACCTTGAAATCGTACATTTTCTCCACCCCATTCTCTAACACAATTCTCA
    TTTTGAGAAGAGGTTCAAGAATATTTATGGTTTAAAAATCTGGAATAAGCATAGATTATACACAACAATA
    TTTTATGTATTTTATTTTTCCCCCTTTGTTCTTATTTATTTATTTATTTATTTATTTACTTTTATAGAGA
    TGGGGTCTCACTATGTTGACCAGGCTGGTCTCAAACTCCTGACCTCAAGAGATCCTCCCATCCCGTTCTC
    CCAAAGTACTGGGATTACAGGCGTAAGCCATCATAGCTGGCCCACACCAATATTTTAAATAACGCAAAGA
    AAATAGGCTGAAAAAAAAGACAACACATATTTGATAAAGGGTGAGAAAAAAACCATTATTGTTTTCCCTC
    CCTCAACTTTCTTTTAATTAATATTAGATGGAAATGGTCTCTCGTTACTTGAAAGTATTCCTGATATTCC
    CAGCTCAGGAATGGAAAACCAAACATCATATCTTCTCACTCAGAAGTAGGAGCTAAGCTATTAGGAAGTG
    GCAGCTAAGCTATTAGGAAGTGGGAGCTAAGTTATGAGGATGCAAAGGCATAAGAATGATACAATGGACT
    TTGTGGATTCAGGGGGAAGGGTGGAAGGGGGTGAGGGATAAAAGACTGCAAATTGGGTACAATGTATACT
    GCTCGGGTGATGGGTGCACCAAAATCTCGGAAATCACTACCAAAGAATTTATTCATTCATGTGACCAAAC
    ACCACCTGTTCCCCCAAAAACCTATGGAAAAAAAGGGAAAGTATTCTGATACTTCCGTGGTGTAAGGCTG
    AATGGGTCAGAATGGGGCAGCAATGCAGAGATGATGGAAATACTCACTGTCTTCCTGCTTCACAAGCAGT
    CCATGAGTCCCAGTTCGGACAATGCCTCCCTTTGCCATTATCTGCAAAAAAGGAAATAAAAAGAAATTAA
    ATGTGCCAGAGAGAACACGCTTCAGTCTCACTTCAATATACTTGTTTTATTGAGTCCCTGCCGGCAATAA
    TTCACCTCAAGAGAAATCAAACTTTCAAAAGGAACATGGATAAGAAGTTGAAGTAATGACATCTAAAGAG
    CTTCATGATTACTTGCTCCACAGAGAGCTTTTGTCTACAGTGAAGTCAACGCAGTAACCTCCCTTCTCAC
    CAGATAATAGAAGGAGAGCTTCTTCAGCCCCAGCAGGGTGCCTCCATTCAGAATATAATGTGCCTGGACT
    GTCTGAGTATGGCCACAGGGTAGTTCATGAGACATGGGCTCAAGGTGGACAAAGCTCTTGCTTGGGGAGA
    ACACAAGATAAGCAGTGTGATGTGCCTCTTCGTGTTCTTCTGACACCCACTGGTAGCCGTAACAGGGACT
    ACGATCCTTGTAATTGACCTAATGAATTAGAAAAATTATATTATTTTTAGATTATACGTCATAAAGATTA
    ATGTTCTCACAAAACTACGTAAGTTTACTGTCCTACCTTAACTAGTAGCTAGAAATCTTTTGTTCCACAA
    TGAAAAGTGGGAGAAGGACCCCTGATTTTGTTTCTCTGCTTCAAATGTAGGCATGCTTTGTGACTTTACA
    CTGCAGAGCCTCCTGGGATTGAGATTTATATTTCTTAAGAGCATAAAAGGAGGCATCCTTTGCTCTGATT
    AGCTTGCTTGAAATTTCTTTGACTGCTTTACCTGTGCAATTAGAGACACTGGAGCTATGAAACTTCATTG
    AGGAGAGTAAATAATCTCTAGAAATGGAAGAACTGTGCTTTCAGAAAGCAGCTGCCTCTTTTAGGGTGGA
    TTTTGAATGGGTATTATGAATAAATAAAATATTGTAACCTTAGGGGTCTGAGAAGACATTGGTAGAAAGC
    ATTGGTTTAAGAGCTGAGCTGATGAATTAGCTGAAGTCTACCTCTTCTGGGCTATCTTAGGCTGATTGCT
    ATTATTATTATTTTTTTGAGACAGGGTTTTGCTCTGGCGCCCATGCTGGAGTACAGTGGCACAATCTTGG
    CTCAGTGCAATCTCTGCCTCTGGGGCTCAAGTGATCCTCCCAGTCAGCCTCCTGAGTTGCTAGGACTACA
    GGTGCATGCCACCCCACCCAGCTAATTTTTTGTATATTTTGTAGAGACAGGGTTTCACCACGTTGCCTAG
    GCTCGTCTCGATCTCCTGGCCTCACGCAATCCACCCACCTCAGCCTCCCAAAGTGCTGGGATTACAGGCG
    TGAGCTACCGTGCCTGGCCTAGGCTGACTATTTTTAATGCTCCCTGCAATCTGGATTCTAATACCTCAGA
    GGCCCTATTTGGGTTATATTTCCAACTTTTCTTCCTGTAGAAGTTATTCCTTAGCTCCTTTCAGGCTTCC
    TCTACCTTCATCCCCAGCTCCCAATAGCACTAGTGAATCCTTGAGCAAAACATTTTGTTCATATCTTAAT
    GTATATTTTGGTGACAACACATATATGATCACCCCAAACAGGGAACAGGTAGAAATAATAATTTCATTGT
    CAAGACTAGCACAGTGCTGGCACGTGGAAAGCATTCAAGACACTTTTTGAAATGAGTAAACATTCAAATT
    AGAGGGAGGTGCTTTTATGCCATCTATTAAAGCTAAATAAATAAAATGTATTATTTATTTCTCTATAACA
    AGGTAAAACTCTAAAGAGGCAGGCGTTTTCCATTACCATGATTATGTAATATATATTAAATATGTATTGT
    ATATCCATGTATCTATAGACATGTCTACATCTCTCTTGATGAATAGAGGACTGCATGCTATACCGAATAA
    CTTATATTGGTATGAACAGGTCATCTGGAGAAAATTAGAATAGCAAGACAAAATAAAGTAAGGTAATAAT
    TTTTGGAGTATATATATTTAAAACTAAATGCTAATTATGTTAGAATTGGCTTTTATAGTAAAAGTATCTA
    ACAGTTAAAGGGGGCTTACTAAGTGATTCAACATGTATGATTTCACTTAATCTTCACAACCACCCCAAGA
    GTCAGGTGTTAGTATTGCAATCATTTTACAGATAAGGAAATGAAGCACAGAGGTTTTAGTTAATTCCTTA
    AAATAGGGTTGAGCCAGTATTTTTAACCACTTTATCTGATTCAGATTCTGGAATTCACACTTTTCAAAAA
    AGTTATTATAAACACACACACACACACAACATAAAATTACCATCTTCATTATTCTTTAGTGTACAGTTTG
    GTAGTATTAAGTATATCTGTTGTGCAACAGATCTCCAGAACTTTTCATCTTGTAAAACTAAACCTGAATA
    CCAATAAGAGGAATTCCCCATTTCTCTCTTTCCCTAGCCCTTGGCAACCACCATCCTGTTTTCTGTTTCT
    ATGACTTTGACTACTTTAGAAACCCCATGTACGTGAATCATACAGTATTGGTCTTTTTGTGACTAGCTTA
    TTTCACTTACCATAACGTCCTCAAGGATCATTCATGCTGTAGCATGTGACAGGATTTTCTTCCCTTCTCA
    GGCTGCATAATATTCTGTTGTATTTATATACCACATTTTGTTTACCTGTTCATCCCTTGATGGACATTTG
    GGTAGCCTCGATCTCTCAGCTATTGTGAATAGTGCGGCTATGAACACGGGTGTGCAAATAGCTCTTTGAG
    ACCCCGTTTTCGATTCTTTTGAATATATGTCCAGAAGTAGGATTACTGGATCATTTGGTAATTCTATTTT
    TTAATTTTTTCAGGAACTACTATACTGTTTTTCATAGTGGCTCCGCCATTTTACGTTCCCAGCAACACTG
    CTAAAGTCTAACCCGCACTTTTAATCACAATATACTGCCTTCTTTTCATAAACTTTAAAAAAGTTAACCT
    TCAATCGGTTTCTAATTGCTAGTTTTTTTCTCTCCATAGAACTAGACACAGTTTGGAAATTTTCTCACCC
    TTAGGTTGCAACCTTGTTTCCAAGGCTACTTCATGTCATTGGTAATTTCTTTCCAAACTTACCCTAACAG
    TAAGAGAGGTACCCATAACATTGGTGGTGTTGATAGAGAACTGTACAAGGCCATGCTCATCCGTGGTAGC
    ATTGGAGTAATAGTTTGCTTCATTTCCTCTGATGAATATGACTTTATTTGGTATAGGGACGCCTTTCCCA
    TCTACTAGGCGCACCTGAAGATTAAAAGCTGTGGATTAGTTATATTGGTAATAACTAATAGGCACCAAAC
    ATATTCATTTATCACATTTTTTAGTGCCTCCTCTATGTTGAACATGGTTCCAGGCACTGAGGACACAGCA
    GTGAAAAAAAACGAAGTTTCTTCCATCACAGAGCTTACTGTCTAGTGAAAGAGATAGATGCTAAATAAAT
    ATATACATAACATACACTAATTTAGTAGTAATCAATCCTAAGAAGAAAAACAGAGCAGTGTGAAGGGATA
    GAGAGTGATAGGGATAATAACAAGAGTGACCTGAAAAATGTCTTCTAGATATTTTGGAAGTGCTGTGTAT
    TATTATATATAACTCTTAGAGAGTCACAGAGCACTAGAGTAATATGATAATGAAATATTTTAATCTATTA
    ATCTATTAACTAAATTTATTTGTGCCCCAAACACCTATTTTGGAGTGGACCATTAACTTCCTATAGATTA
    GTGTCTGAAGTCTCATAAGTGCACAAGTAAACAATTGCCAATACATTGTCCTGTGTGTTTCTCCAGTTGC
    CTTAGACATGGTCTCTGCTTCTCAACCTAGATCTTCAGCCTTTTGGAAGGTAGGCAGATTTTATGACACA
    CTTCCCATGTGAAAAATATAAAATTGAAAAGAGGCACACAGTATTAAATAAGTTGCCAAATCTAATTTAC
    TGCTAATATTGTATTGGCAGTTGAGTAGTCCAGTTCAGATTGTAAAGGTGGCAATTCTTTTTCAGTATAA
    GCTACAGTTGGTAGCAGGAAGAACAATAGGCTGACCCCCATCAGTACTCATAATTGATTTACAAAATCTA
    AGTTTGCCAGTGAAATGTTGGCTGCATGTACAAAGACAAGAATAGGCACATAATTCTGATTTTCTTCTAG
    CTAGAGAGTCTTATGACAAATAGGATATTTAACAGCAATAAAATGTACTTACAAAGTACACTATCTTTCT
    GAGAAATTTACAAATGAGCTACACTTTCATTTTAGGAAGTTGAATACATTTTGGTGTCATGACATAACCA
    AGCCTTCTAGAATGTACTCCACTTTGTTGAGTCTATGCCTCTTTTACACCCTTCATAGCTGAAATCCTGT
    AACATTTACATTAAGAAAATGATACACAGTAAATACACATTGAGGCACATGTAACTGTGATCTCAGACTT
    CTAGAGTCATAAAGATCCATTTGCACATCTGCTGACAACAGAAAAATTGTAAGAGAAGATTTTTTTGGTA
    GATAACATGAAAATTCATAATTCAAAAATCTGAGAACCTTAAATAGCAGTGTTTTACCCCATGTTCACTG
    TTTTAGAAATATACTTGTTATAGAAAAATAGTGTAAATATACTATCCATGAAATAAACATTATTGTAGAA
    TCTTTAATACTTCAAAATAAAAATTAAAACCACCTACCCACCTGTTTCCATACAAAGTTAGTGTTAGCAT
    CGTAATGAAACTGGTCAGCATGAATTTTGAGATTCTTGTGAGAAAATGTCATAAAAACCTGAGACTGCCT
    GGTAAACCATGAGAAGGAGATTGAGTGACTTGTACAATGCTGTTTCAAAAGAAGGGGGGAAAACTCTACT
    AATTTTAATAAGGCCTTAGGATAATTACATACATTTCATACACAAATTTACAGAAGAATAAATCTATTTT
    TATGTAGTTTTATACCTTTGGTTATACTAATCAAGCCTTGGTGATATTTTATACTTTTGGTTATATTAAA
    TGATAACATTAACCAGGCATGGTTTTAGCACAAACCATAACTATTGTGCTAATTAGGTAACAGTACATGG
    GTTGATGAGTGAACTGGAAAATACTCCACCTGCCCAAAGAAGGGAATTCCCTGTCGAAAGTGTGAGTCCA
    CTTTCACAAATGAGAGTTTGGTTATGGTTCTTGTGATTTCACTGGACTGCCTTCCAGTCAATTCCACCAC
    TGAAAAAAGAGAAAAAAATCTGTTATTTTTGGGAAGAATGATGTCTCTAAAATTCTATCACCTCCCCCCA
    ACTTACAATTCATTATTTGGCTAAGAAAATCAATGCCTGTTGTGTTTTCTCTTATACCCATGTAGTACAC
    AAACCTGTTCCTTCTTCTTGGATCTGGGCCTCAGTGTGAAGTTTCATTTCATACTCCTTCCTCTTCAGCT
    GGAAGACCTTGGTTTTTACTTGCTGATAGAAGCAGCCATGGCTGTTTAGCTAAGAAGGGAGAAAATAAAA
    TACAAAAATATAATGCATATTATACTAAATAGATCAGTGGTCAGATAACCGGTGTGATTTTTGTGGGGGG
    ACAACATCATGTAGGCATTTTCTATGACGAGGACACAATAAATATTCTCTAGCAACTAAATATTAAATTT
    CACAACAATATCTCATGTATAATTGACAGATGTATTCCTTGGTATCTGAAATTCCCTAGATATATTGCGA
    TAGAAATGTCTTAAAATTTGAGGAGTAATTTTTTTTGTACAAAGGATGCATAATATTTATACATATTTAT
    GGGGTACACGAGATATTTTGTTACATGCAGAGAATGTGTAATGATCACATCGCGGGATTTAGGGTGCCTA
    CCAACTCAAGTATGTATCATTTCTATGTGTTGGAAATAGTTCAAGTCCTGTCCACTTGCTATTTTGAAAT
    ACACAGTACATCTGCTCTTGACCATTAGAACTCCTCATCAAAGTCTGGTTCCTCCTCCATAATATTCAGC
    TGTGGCTGGGAAGCGGCCTTCTATATGGGGACTGTGTTTGTCAACTCTCTGGCATCTTGGTGGGGCCATG
    TGATAGGTTCTTGTTCACAGAGAGAGCAGAACTGATGGCCTGGCCAGCTAAGGCAATTAAGAGGTGTATG
    GTTTTTTCCTTCCTTTGTAGAAGTGAATGCCCATGGAGGCTTTAGATGGCTCAGTGTCTATCGTTCTGAA
    ACACTCACAATGGGCAATTACGAGAAATAAATAATTCCCATTGTGCTAAGTTCATGATTTTTTTTGAAAA
    ATTACAATAGCCAACATGGAATTCTCTTCTAGATTGTTCTCTTCCTGCGTCAGAAAAATGCTGCAACTCA
    CTGCTTTTCAACAATGCCTTTATCGCTATTCTCTAGAAAAAATAGTGTTCAACCTACCTGTCCACTGAAT
    TTCTCACAGAAAGCCTGTGAATCTTCACCGTGGCAGTCGGAAGCGTCACTATACTTTCTGCAAATGCTCA
    CAGTCACATGTCCAGGGACAGGCTTCCCATATGTGTATCTGTCATGAGAACATTCCCAAAGGAATCAGAG
    CAGACACTGAACCTCCCCATTTTCTAGCTATTTATATCTCCCCTTTAGCTACAGTATATTCCCTGTCTGT
    ACTTCCCTCTGCTCTCTGCTGGGCTCACACAATATTGAAGACAAGAGTCACTAATGGCTCTTATAGAAAT
    CTCCATTGCAACATGCTGAAATTTACATTCTTCTTTTATCATTTCTTTTTTTTCCTCGTTTTTTTTTAGT
    TTTATCTTAAAAAAGTCTTTCTTATTATAAAACTTAAACCCGTTCTTAAAAGAGAAACAATACCAGCAAG
    ACGAAACAAACAAAACAAAACAGAACAAAAAAAACCCAAACGCCCTCAAGTCTACCTAGGAATACAAACT
    ATTAGTACGATAGTGTCTCTTCTTCCATCATCCTCTCCCCATGTGCAGCCATCTCTATATCGAGTTTCAC
    TTGTTTATTTTATTTTATTTTATTTTATTTTATTTTATTTTATTTTTGAGATGGAGTCTCGCTCTGTCGC
    CCAGGCTGGAGTGCAGTGGTGCGATCTTGGCTCACTGCAACCTCCACCTCCTGGGTTCATGCCATTCTCC
    TGCCTCAGCTTCCCAAGTAGCTGGGACTACAGGCGCTGGCCACCATGCCTGGCTAATTTTTTGTATTTTT
    TAGTAGAGACGGGGTTTCACCATGTTAGCCAGGATGGTCTTGATCTCCTGACCTCGTGATCTGCCCACCT
    CAGCCTCCCAAAGTGCTGGGATTACAGGCGTGAGCCACCACGCCGGGCCTGTTTACTTTTTTACAAAAGT
    GGGATTGTGTCGTAAATATGTCATAGACAATTGCTGGGTTTCCAAGTGCATTTGTCTCCTTTCCTATTTG
    AGAATGTCCTCTGTATGTCTATTTGCAAGATCACACCAGTTATGAGGAACCAGGAACTTGGCCATCACTG
    TGAGACAGAACTGGAGCAGGGGCCTAGCTGAATAGTAGGTGTATTACATGTGTGATCTTTGGACAGGACT
    ATACCTGCTCTTCCTGCTACTGCTACTGTGTGCCACCATGGATGGGAACATAAGATCCAGGAAACAAGAC
    TTTTCAAGGTTGGAACAGACTGTCTTGAGAAATCACGTTCAGTCTCTTCGTAAAAATGTCAGTGCAGACA
    GGTATGTAGACCCGCCTTAGGTGGACTAGTGAGTTAGATGAGATTTCAGTTGTATGCCTATCTCTACAAA
    TTCTGAATCTAAGTGATTTTTCCAGTAAGCATGCAAGGTCAGGTAAATGATCTGTTGATGATTGAATTTC
    CCCTATAGCCATGTGTTAATTATCTCCCTAATATACGTTTGGTTATTTTTGGTTAAGGCAGAACCAATAC
    CCTTCAACACTATCCACCCTTCCCATTTTGTGTGTGTGTGTGTGTGCAGTTTTGTCACTTTCAGAATCAA
    TCCTATTAGCATAACCTCTCTAGAAACAGTTTCTTACCTCATGATAGAATAATACTGATAAGAACGTGAA
    AGAAAATAAAAATAGAAAAGCAACAGGAGAATAACATTCTACAGATGAGGATGCTGTCAAAGCACTTAAA
    ATTTTTGTGTTGCCACTGCTCCCGGAGAGAGTAGTTAAAATATCCCAAATGGTGAGTCTCTTTTAAATAA
    TCCCCCACAAAAGATTTTTAAAAAATGAACTCACAGGCCACACACTGATACATTCATCTCTTCTTCCAAG
    ATGGTGATTATCTTTGGCACTGTTACTTGTACTTCAAACTTGGGAAGAACTGTTGATTGGTGATAAAGAA
    GGTTGGTGATTGGTTTTCAACTTTGGGGGAATTCCTATTTTCAGGTTCCCTGTAACAGTTCCCTAATAAT
    ATTTTAGGGCTCTGAAAAGGTAGCTCTGCTCTCCAATCTATTCTTATCTATCCTCCTCTCAAATGAAGAT
    TTCAGTTGAGGGACTATGAGAGATCACTGTGTGCTTAATTAATTATGGAAAACTCCATTAAAACAAGAAT
    GTTTTAATTTTGGGGGTTGGTATTAAATATTGCTACAGAAAATAACGAAGCTATGTGAAGTCCTATTCAC
    AGATCACTGATGTCCTCATTGGACTTAGGTGTAATTAATTCTACTCCAAGCCCAATGTCACCCATGGGAA
    ATGGAAAGACTCCAAATGAACTTAAATTCCAGGACCTAAGAGTTTAAATATGAAGAAAAATAATGCTTGA
    TGACTTTTCATGATCCATACCAAATTCCTCCACGGTGAAAGGGTGCTCTGTCCTTCCACCTGATTTCTTC
    TGTACCACCACCTTGTAGGAGCCCTGGAAGGGCTCTGATGAGAGGGGAAAAGAAAATTGCTTGAGGCCAC
    CCTCTAACTGGAAACTCTGCCATTGTGCGATGCGATTTCCTTTGGGATCCTATATGAGTAAATTAATTAT
    AACTTACAAAAGCACCTGGAGCATTGGAGCTAATAAAAGATATCTATGGAAACCCTAAGTTATCTACAAA
    AAGGTCAAATGGGGTAGTAGTAAAGGGTGAGTAGAGGAGATGAGTTATAGCCATCTAGCTCTATGGGAGT
    TTGAAGGCTTCTCCTTTAATGACAAGTTTCTGAGATATTGTAAAAACCTCTGAAATAAGAACATTTTCCC
    TATATGTATTGCTTTCCTTTTAATATGGAATGTTTCATGTTGCTTACCTGAATGTATACTAGTGGAATCT
    GAAAGACAAAAGAAAAAAGAAGTTTATAATTATTTTCAAATATTCTTAAATCTCATTTATAAGCAAAGCA
    GCTAGTATTTGCTAGCATAACAGGGTGAGTATAGTAAAAAATAATTTAATTGTACATTTAAAAATAACTA
    AAAGTATAATTGGATTTGTTCGTAACACAAAGAATAAATGTTTGAGGGGATGGGTACTCCATTTATTGAT
    ACAGGCATGCAATGCATGATAATAATCATTATCCATGATTGGATAATGATATGGATAATGATAATGATTA
    TCATCCATAATCATTATCATGCACTGCATGCCTGTATCAAAATATCTCATGTAACCCATAAATATATATA
    CCTGCTATGTTTCCACAAAAATTAAAAACAATAAAAAGTTTAAAAAATCTAAATTAGTTTCATATAGAAT
    AATTCCCTCTATAGTGAAATACAGAAATATTCCAGATTAAATTTTATAGAACTTTTCATTAATCTATTTT
    AAGCAATTAATAAATAATGTTAAGAACTTACTAGGCCTTCAGCAACAAATATACATGTAAAGTGCCATTT
    ACTAAAAATGGATTAACATGTCTTAAGTGCTTATAGCATCCTAGTGAAGATCAATCCTTTGCTATGCAAT
    TTTACAGGTTGAGAAATAAACACTTATGTAACTATAAGTAAATATGTTCAGATGTAATCATTAAGATGGG
    TCATAGAATTCTTACGTAGTTCTAAGAATGAGGAGGAATGGACATATTTCATTAATAAATTTAGTTTAAT
    AAATATTACTTTTAAAAATGAATAAACATTGATAAGAAGCTACTATGAATGTTTATTCATTTTTAAAAAC
    TAATCTTTATTAAGTACCTTCTCATGCGCTGTGTATCATTCTAGGTATTGAGAATATAACCACGCACAAA
    AGAACAATAAACCATGTTTTAATGGAGCTTATGTTATAATAGAAATAGGCAGATTTTTAAAAATCAATAC
    GTATAGAATGGTGGTAAGTGCAAATAGAAAAACAAAGCCAAGTTAGGCTGACAGAGTGATGGGTGTGGTG
    CCTCATTTTAGACAGAGGAGTGAAAGTAGAAGCTGGGAGACCACTTTGAAGGTTATTGTAATGATCTTAT
    TTAGAGATGACATTGACTAGGACTAGCTTGCAGCAGTAGTTCAGATGATAAGAAATAGTCTGAATATATT
    TTAAGCCAACATGATTTTCCAGTGAAGTGGATGTGGGTATGAGAGAAAGAGAAACAAGAACTAGGCCATA
    ATCTTTTGTCTTTGGAGCTAGAAGAGAAGAGTTGCCTCTTTTTGAGATAAGAAAATGTGAAGAAGTTAAT
    TTGGGAGAGAAAAATGAAGACTTTGGGTTGATGTCTGCATTACAATAGAAAGAACTGTTTCTTATCACAG
    ATACCAAGAGTTAGTCTTGAGGAACAAATTAATTACCCTGACTTTCCTTTATAGTGAAGATTTTGGTAGA
    TGTGTATAGAAAAAATCTAAATCTTTAAAATGTTTTTTGAGCTTAGGTTCTTTGAGGATTTTTCTGTGTC
    TGCTATTGAATGTATCTTTCTAATTGTCCTAATTAAGAACCATCCGAAAAGTAAATTTAATTGTGTGACA
    ACTGACTGAGTACCAGCACCAAGACAGAAAGAATTACCTACTTTCTTTACCTGGAATGATGTTTTAGATT
    AGGATCTTGTGCTGTTGTAATGCCAGAAGTTACTGTTAATGATACCAGATTCTTCCTCTCCCTGGTCTTC
    CCTCAGCACAAAAAACAGGTTCCCAGCCTGTTTATACAGACAATCATTTTATGTAGATAATAGAAAACTC
    ACCAACTCATTCAGGGGGTGAAAGTTTTCATCCATGGAGACAACACGAAATTTCACTGAAACAGAAATAT
    TTTTCATGAGCCCCCAAACCCAAAGTAGGCCTGTAGGCTGGTAAGACAAGCTATTAAGGAACCAAGATTA
    TAGGAACTTTGCCTGGGGAACATCCAGGGGAAGAAGATCTTTCCTTTTTGAAGTTGTCCTGTCTGTAGGC
    TTCTTCATACCTGTCTGCCCTGGTTTGTAGATTGATTTGTCTGTCTGGACAAAGACCAGACTGTCCTCGT
    TCTTAACCATCACTGTGGTCCGCTTCTTAAATTCTTGGGTTGGTCCTTTCACTTGGACAGTGAGGAACAT
    TACCTCCTCATTGGATGAAGACTTTGGGACCTGAAATACAGGACCGATCCTGAAACCCCATTCAGCACCC
    GCAGGTCTCCTCCCCTATTTGCTGTTGCACTCTTCCCTGGAGATCTTGCCCTTCTTACTTAACTTCACTC
    CCTTGTTCTCAGCAGGGAAAGTTGGCATGGACAGGAATCACTTCTGCCATTTTACAAATGGGGAGACAGG
    ACACAAGGTGGAGGAAAATAAATCAGTAAGAGAGGTTGTAAGTGAGATTCACACCTTCATACAGCTCACA
    GAAACAGTCTTGATTCAATTATACGCTGAGCTTTACCAGGCTGATATTACATGCTAAAAGGGCATGGTCG
    AAGCATGTTTTACAGGCTATGGAAAAAAACCCCGTATTTTACTGACACTAGGCTGTAAGCATCTCACATG
    TGGACAACTCTGAAGAGATATTACACCTTTGCTCAGGGTAATTTCTACAATTTTTACTCTTAGAAGTAGT
    GTGGAGTGATGTGTTCCTGTCAGTCCACCACGGGTGGCTCATGTTGAGCCTTGGTCTAGTTTGCTTTATT
    AATATTTTTTCCATCTTAATTTTAAGAAGTCACATTTTCTTTTTTTTTTTTTCCTTTCTGGCTAATAGGG
    TTTCTATGTGATTATTAAGCTGACTTTTAGTTTGCCTGTAAATATTTGTCTCCCATGGCTGGAGAGTCAT
    GCATTGCTCCTTAATTTCTATACTTAGTTTGCTACATTTGGATTCCTTCCTGCAGTTCTTACCAACCTCC
    CAAAGCCTCATCTGACAGAATACTAGATCCCTATGACCCTGACTAAAAGAACCAAGTATATTAAGTCAAA
    CAGCCACACTCACAGCGAAGGCGACACAGTGGAGTACGTCATTCTCCGCCTCCAGGTCAGTGAAGAGGCT
    CCTGTTTCCCCTGACAGACTCCAAGGAAGCACTTACAGTCACTGTCTCATTCAGGTAGCTCAGAAGGACA
    CAGCCCTTCTCAGTGGTCTCAGTGTGGAGCAGGGAGGGGACCAGAACCATATACTGCCTGGGAATGAGAC
    GGTTCAGTTAGAGGAAAGTGAAGGAAGAGAGGCATTGCTATCAACAGCACTTTTTTCCATCATCATAATT
    ATCATCATCAGTTCTACACCAAGAGAAGATGTACTGATGAGCATGAAATTTGGCCGTTGAAGGCCATGCA
    TAAGAATTATTTCCCTTTTCCCTCACATGCCATGGCAGTGAACATTTTACTTGAGGTAGGTCTTAAACAG
    ATTAAAACTTGTAGGCATGCTTCACTTTAAGGAACAATTTGTGAACATCTGAATTCAAATAAAAATATGT
    AGTTACATTTGTTCATGTTGTAAAATCTATTTTTACCTTCAAGTTGCTGTAAAAAGTAAGCAAGCTCTCA
    GTCATGCGGACATTACAAAAACTAATCAAATTATAGAATTTTCATTAAAATATATATACACACATACATA
    CATACACACATTCAGCAGCCCTGTGAAAATTGTATGGTAAGCACTCGCTTCCACTCCACAGATATGGAAT
    CTGAGCCTCAAAGAAAGTGAGCCATTACTTCACAGTCTCAGTTGTGGTAAGCAGCTAATAAGGTCTTAAG
    AGTGTTACCTAACCATGGATGTTTGCATTTTAAATATTTGCCTTTCAGCTCATGATTAGACAATGGCTCC
    ATCCAAAAATGGTCCTCTGGTGATTGACTTTAGACCTTGGTAACTCATGTGCAGTACAGAAACTTTCTCA
    GTACCCTGACATGTAATTTAAGATCAACCCTGTAGTTTTTATACATTTTTGCTCACAGAACTCCTAAAAT
    AATTTTTGGACTCTGTGCACTCCTTACATATTTTCAAGTAAACATATTTTTATCATGATTTTAAGTAATA
    GCAAAAGATATTATTTTTGATGATATTTTAAATTAATACTTAGATTACTTTTACAATTTTCTCATGAATA
    ATAAACACTGTAGTATAAGATACCTACTATCACTATTTAAAAGTACATTAAAAAATTCTTTTAATGGTCT
    GATATTTAATATCTTTCTCTTCTTTTTTCCTTTTTGAATTTGAATTTCCATGCCAATTACCATACAGAAA
    TTTATCCTAAATTTTTATATTTCAAAAAATGTTGTGAATTATTCTTACTTCTTTATTGTCAAGCATCTAT
    ACATATACGTATGTATACTTATGTTCACATATATACATATATGAATACATATACATAAATACATATATAT
    GTAACATATGTGGGTAGGTAAATTTAACTTTAACTCCTCTTTTAAATAGAATGTAACCATAAATTTATAA
    ATATTAAACATTTCCTTCTGACTGAGCTATGTTCAAAAATACTTGTATATAACTGATCAAAACAAAATAA
    ATACAAAGTATGATTCATACATATTAGATATAAAATTAAAATACTTTTGAAGCACATCTTTAATGAGAAT
    GATGAGCTGTTTTTTCCCCCAGTGAATTTATGCATCCATGGATTGCAAGAATTAAACAACATTAATTCTG
    TTACTATTTTTGTTTTGTGTAGCTATAAGTGCTAAGAAGAATGGCTCACATAAAAGTAAATAAATGGCAG
    GTAAAACAATTGCATCATTGTTACCACTTCTTGCAACTTCTGCATTACATGCCAAAAATCACCTAGTACT
    CTATTATCAAGAATCCCTTCTCATCACTCATCAGCTGGTATTTCCTTCAGTTCCTCCTTCAGTTGTGTTG
    AAACAAAGAATTCATCACCACCTCATCTGCAAAAGGATTTCTCTTCTGACATTAGAGTCCCTTGCTTTTT
    CAATATCAACACCAGTATTTCAAATTGTTCCTTTATTTGGCACTCTATAGATTTTTATACTCCAGGGCAT
    TGTTCCATGTTTGGTTGTGGGAGAGAATTGTCTTTATTTTTATAAAATAGAAAATGGACCCTTGAGAAAG
    AGGAAATAAGAAAAGATAAATAGCATGGTCTCAAGTAGATCAATATGAATGTGAATATCTAAAATTTATT
    CTTTTAAAAAGCTAAAAATTTCTGACACCCCTTAGGAAGGCTTTGCATATTAGAGCTTCAGAGAGTTGGG
    GAAGAATGATTAAAAGTAAGCAAGTAATGAGTAAACTAGATTTGTAAAATCTGGAAGGAATCTTAGAGAT
    AAGAAGATGACAGTATCATAGGAAAGAAAAAGGAATGATATAAAGAAAAATCTGCAATAAATGAAGGACT
    CTAGGTTCATGCTTCACGCTCTCTGTGTGGAACTCACGGTTTTCCAGAGACTGAGGCGTCTGTGGGCAGG
    AGGACCAAGAGGAGAAGAACCAGACTTGGATGAAGGAGTTTGTTCTTCCCCATGTTGCAGAAAGAAGGAG
    CTGGAGGAGAACAGACTGTATTGTACCCTACTCCCTACAATCCATCTGGTCCCAAACACTTCCCAAAGCA
    ACTGGGCTTTATGCTGCTCTGTGTG
  • Protein Sequence : Show Sequence
    >gi|66932947|ref|NP_000005.2| alpha-2-macroglobulin precursor [Homo sapiens]
    MGKNKLLHPSLVLLLLVLLPTDASVSGKPQYMVLVPSLLHTETTEKGCVLLSYLNETVTVSASLESVRGN
    RSLFTDLEAENDVLHCVAFAVPKSSSNEEVMFLTVQVKGPTQEFKKRTTVMVKNEDSLVFVQTDKSIYKP
    GQTVKFRVVSMDENFHPLNELIPLVYIQDPKGNRIAQWQSFQLEGGLKQFSFPLSSEPFQGSYKVVVQKK
    SGGRTEHPFTVEEFVLPKFEVQVTVPKIITILEEEMNVSVCGLYTYGKPVPGHVTVSICRKYSDASDCHG
    EDSQAFCEKFSGQLNSHGCFYQQVKTKVFQLKRKEYEMKLHTEAQIQEEGTVVELTGRQSSEITRTITKL
    SFVKVDSHFRQGIPFFGQVRLVDGKGVPIPNKVIFIRGNEANYYSNATTDEHGLVQFSINTTNVMGTSLT
    VRVNYKDRSPCYGYQWVSEEHEEAHHTAYLVFSPSKSFVHLEPMSHELPCGHTQTVQAHYILNGGTLLGL
    KKLSFYYLIMAKGGIVRTGTHGLLVKQEDMKGHFSISIPVKSDIAPVARLLIYAVLPTGDVIGDSAKYDV
    ENCLANKVDLSFSPSQSLPASHAHLRVTAAPQSVCALRAVDQSVLLMKPDAELSASSVYNLLPEKDLTGF
    PGPLNDQDDEDCINRHNVYINGITYTPVSSTNEKDMYSFLEDMGLKAFTNSKIRKPKMCPQLQQYEMHGP
    EGLRVGFYESDVMGRGHARLVHVEEPHTETVRKYFPETWIWDLVVVNSAGVAEVGVTVPDTITEWKAGAF
    CLSEDAGLGISSTASLRAFQPFFVELTMPYSVIRGEAFTLKATVLNYLPKCIRVSVQLEASPAFLAVPVE
    KEQAPHCICANGRQTVSWAVTPKSLGNVNFTVSAEALESQELCGTEVPSVPEHGRKDTVIKPLLVEPEGL
    EKETTFNSLLCPSGGEVSEELSLKLPPNVVEESARASVSVLGDILGSAMQNTQNLLQMPYGCGEQNMVLF
    APNIYVLDYLNETQQLTPEIKSKAIGYLNTGYQRQLNYKHYDGSYSTFGERYGRNQGNTWLTAFVLKTFA
    QARAYIFIDEAHITQALIWLSQRQKDNGCFRSSGSLLNNAIKGGVEDEVTLSAYITIALLEIPLTVTHPV
    VRNALFCLESAWKTAQEGDHGSHVYTKALLAYAFALAGNQDKRKEVLKSLNEEAVKKDNSVHWERPQKPK
    APVGHFYEPQAPSAEVEMTSYVLLAYLTAQPAPTSEDLTSATNIVKWITKQQNAQGGFSSTQDTVVALHA
    LSKYGAATFTRTGKAAQVTIQSSGTFSSKFQVDNNNRLLLQQVSLPELPGEYSMKVTGEGCVYLQTSLKY
    NILPEKEEFPFALGVQTLPQTCDEPKAHTSFQISLSVSYTGSRSASNMAIVDVKMVSGFIPLKPTVKMLE
    RSNHVSRTEVSSNHVLIYLDKVSNQTLSLFFTVLQDVPVRDLKPAIVKVYDYYETDEFAIAEYNAPCSKD
    LGNA
  • Molecule Role : Vaximmutor
  • Related Vaccine(s): Actinobacillus pleuropneumoniae HS93Tox-/pIG-T1K vectored vaccine
2. Ighg1
3. Ighv1-9
  • Gene Name : Ighv1-9
  • Sequence Strain (Species/Organism) : Mus musculus
  • NCBI Gene ID : 668478
  • Genbank Accession : AC073561
  • Taxonomy ID : 10090
  • Chromosome No : 12
  • Gene Starting Position : 114583568
  • Gene Ending Position : 114583861
  • Protein Name : immunoglobulin heavy variable V1-9
  • Protein Note : Also known as Igg2a; Gm16697
  • DNA Sequence : Show Sequence
    >gi|372099098:114583568-114583861 Mus musculus strain C57BL/6J chromosome 12, GRCm38 C57BL/6J
    GTCTTGCACAGTAATAGATGGCAGAGTCCTCAGTTGTCAGGCTGCTGAGTTGCATGTAGGCTGTGTTGGA
    GGATGTATCTGCAGTGAATGTGGCCTTGCCCTTGAACTTCTCATTGTAGTTAGTACTACCACTTCCAGGT
    AAAATCTCTCCAATCCACTCAAGGCCATGTCCAGGCCTCTGCTTTACCCACTCTATCCAGTAGCCAGTGA
    ATGTGTAGCCAGTAGCCTTGCAGGAAAGCTTCACTGAGGCCCCAGGCTTCATCAGCTCAGCTCCAGACTG
    CTGCAGCTGAACCT
  • Molecule Role : Vaximmutor
  • Related Vaccine(s): A. pleuropneumoniae DNA vaccine pcDNA-apxIA/pcDNA-apxIIA
IV. Vaccine Information
1. A. pleuropneumoniae ApxIa and ApxIIa protein vaccine
a. Vaccine Ontology ID:
VO_0011384
b. Type:
Subunit vaccine
c. Status:
Research
d. Antigen
A. pleuropneumoniae ApxIa and ApxIIa
e. Gene Engineering of ApxIIA
  • Type: Recombinant protein preparation
  • Description: The apxIIA gene was cloned from A. pleuropneumoniae serotype 5 isolated from the lungs of Korean pigs with pleuropneumonia. For the oral vaccine, S. cerevisiae expressing ApxIA antigen as well as the ApxIIA antigen were prepared (Shin et al., 2007).
  • Detailed Gene Information: Click here.
f. Gene Engineering of ApxIA
  • Type: Recombinant protein preparation
  • Description: The apxIA gene was cloned from A. pleuropneumoniae serotype 5 isolated from the lungs of Korean pigs with pleuropneumonia. For the oral vaccine, S. cerevisiae expressing ApxIA antigen as well as the ApxIIA antigen were prepared (Shin et al., 2007).
  • Detailed Gene Information: Click here.
g. Adjuvant: Freunds emulsified oil adjuvant
h. Adjuvant: complete Freunds adjuvant
i. Vector:
Saccharomyces cerevisiae
j. Immunization Route
Oral immunization
k. Mouse Response
  • Host Strain: BALB/c
  • Vaccination Protocol: Briefly, 15 mice per group were subcutaneously injected with 100 µg of protein extract after emulsifying with complete Freund's adjuvant (Sigma, USA). This was then followed by a boost immunization with the same amount of antigens after emulsifying with incomplete Freund's adjuvant (Sigma, USA) at 2 weeks after the initial immunization. The final immunization was performed in the same manner at 2 weeks after the boost immunization. All groups were immunized orally through an oral gavage with 4 doses of Saccharomyces cerevisiae expressing either ApxIA (group C) or ApxIIA (group D) alone or both (group E) at 10-day intervals (Shin et al., 2007).
  • Challenge Protocol: Mice in each group were challenged by intraperitoneal injection of a field isolate of A. pleuropneumoniae serotype 5 at 1.45 × 106 CFU (minimal lethal dose, MLD) in 10 days after their final immunization, and were then monitored every 6 h for up to 72 h (Shin et al., 2007).
  • Efficacy: After the challenge, the mice in group E had a significantly lower infectious burden and a higher level of protection than the mice in the other groups (p < 0.05) (Shin et al., 2007).
2. A. pleuropneumoniae ApxIA protein vaccine
a. Vaccine Ontology ID:
VO_0011356
b. Type:
Subunit vaccine
c. Status:
Research
d. Antigen
ApxIA (Shin et al., 2007)
e. Gene Engineering of ApxIA
  • Type: Recombinant protein preparation
  • Description: The apxIA gene was cloned from A. pleuropneumoniae serotype 5 isolated from the lungs of Korean pigs with pleuropneumonia. For the oral vaccine, S. cerevisiae expressing ApxIA antigen were prepared (Shin et al., 2007)
  • Detailed Gene Information: Click here.
f. Adjuvant: complete Freunds adjuvant
g. Vector:
Saccharomyces cerevisiae (Shin et al., 2007)
h. Immunization Route
Oral immunization
i. Mouse Response
  • Host Strain: BALB/c
  • Vaccination Protocol: Briefly, 15 mice per group were subcutaneously injected with 100 µg of protein extract after emulsifying with complete Freund's adjuvant (Sigma, USA). This was then followed by a boost immunization with the same amount of antigens after emulsifying with incomplete Freund's adjuvant (Sigma, USA) at 2 weeks after the initial immunization. The final immunization was performed in the same manner at 2 weeks after the boost immunization. All groups were immunized orally through an oral gavage with 4 doses of Saccharomyces cerevisiae expressing either ApxIA (group C) or ApxIIA (group D) alone or both (group E) at 10-day intervals (Shin et al., 2007).
  • Challenge Protocol: Mice in each group were challenged by intraperitoneal injection of a field isolate of A. pleuropneumoniae serotype 5 at 1.45 × 106 CFU (minimal lethal dose, MLD) in 10 days after their final immunization, and were then monitored every 6 h for up to 72 h (Shin et al., 2007).
  • Efficacy: The immunogenicity of the rApxIA antigen derived from the yeast was confirmed by a high survival rate and an ApxIA-specific IgG antibody response (p <0.01) (Shin et al., 2007).
3. A. pleuropneumoniae DNA vaccine pcDNA-apxIA/pcDNA-apxIIA
a. Vaccine Ontology ID:
VO_0004544
b. Type:
DNA vaccine
c. Status:
Research
d. Host Species as Laboratory Animal Model:
Mouse
e. Gene Engineering of apxIA
  • Type: DNA vaccine construction
  • Description:
  • Detailed Gene Information: Click here.
f. Gene Engineering of apxIIA
  • Type: DNA vaccine construction
  • Description:
  • Detailed Gene Information: Click here.
g. Vector:
pcDNA3.1 (Chiang et al., 2009)
h. Immunization Route
Intramuscular injection (i.m.)
i. Mouse Response
  • Immune Response: Significant humoral immune responses were induced by this DNA vaccine through intramuscular immunization. The IgG subclass (IgG1 and IgG2a) analysis indicates that divalent DNA vaccine induces both Th1 and Th2 immune responses (Chiang et al., 2009).
  • Efficacy: Animals immunized with divalent vaccine demonstrated 70% survival after challenge with a dose of 5 × 108 CFU of A. pleuropneumoniae serotype 1 ten days after 2nd boost. Survival was significantly higher than that of the negative control groups (P < 0.05). No protective efficacy was observed for pcDNA3.1 vector immunization group and PBS control group and all mice were died within 24 h (Chiang et al., 2009).
  • Host Gene Response of Ighg1
    • Gene Response: Both IgG1 and IgG2a responses were induced by divalent DNA vaccine, with IgG1 slightly higher than IgG2a. The levels of IgG1 and IgG2a were also significant compared with the negative control groups (Chiang et al., 2009).
    • Detailed Gene Information: Click here.
  • Host Gene Response of Ighv1-9
    • Gene Response: Both IgG1 and IgG2a responses were induced by divalent DNA vaccine, with IgG1 slightly higher than IgG2a. The levels of IgG1 and IgG2a were also significant compared with the negative control groups (Chiang et al., 2009).
    • Detailed Gene Information: Click here.
4. A. pleuropneumoniae HS93C-Ampr
a. Vaccine Ontology ID:
VO_0011385
b. Type:
Live, attenuated vaccine
c. Status:
Research
d. Antigen
A. pleuropneumoniae ApxII-activating protein, ApxIIC
e. Gene Engineering of ApxIIC
  • Type: Gene mutation
  • Description: Site-specific mutagenesis of the apxIIC gene utilized the recombination plasmid pEP-CAmpr. Cesium chloride-purified pEP-CAmpr DNA was isolated from E. coli and linearized with ClaI. Following digestion, the DNA was purified by phenol-chloroform extraction and ethanol precipitated. A total of 3 μg of linearized DNA was electroporated (0.2-cm-diameter cuvettes; 400 Ω; 1.25 kV) into A. pleuropneumoniae HS93 (serovar 7, ApxII) (Prideaux et al., 1999).
  • Detailed Gene Information: Click here.
f. Vector:
A. pleuropneumoniae serovar 7 strain HS93
g. Immunization Route
Intramuscular injection (i.m.)
h. Pig Response
  • Vaccination Protocol: Nine 6-week-old pigs received 109 CFU of the A. pleuropneumoniae vaccine strain in 1 ml of growth medium, via intranasal inoculation on day 0, while nine control pigs received 1 ml of BHI. The vaccine was prepared by inoculating 10 ml of BHI-NAD (10 μg/ml) with a single colony of the vaccine strain and growing with shaking at 37°C until an OD600 of 0.8 was reached. The vaccination schedule was repeated on day 14. On day 28, the nine vaccinated and nine control pigs were divided into groups of six and three (Prideaux et al., 1999).
  • Challenge Protocol: Two groups of six pigs (i.e., vaccinated and unvaccinated) were challenged with 2 × 109 A. pleuropneumoniae HS25 (serovar 1) in 2 ml of growth medium via the intranasal route, while the groups of three were given 2 ml of BHI broth in a similar manner. The challenge strain was prepared by inoculating a single colony of HS25 into BHI-NAD (10 μg/ml) and growing until an OD600 of 0.8 was reached. At this time the viable count was 109 CFU/ml. At 5 days postchallenge, pigs were euthanized, and the number and severity of lung lesions were recorded (Prideaux et al., 1999).
  • Efficacy: Pigs vaccinated with live HS93C- Ampr via the intranasal route were protected against a cross-serovar challenge with a virulent serovar 1 strain of A. pleuropneumoniae (Prideaux et al., 1999).
5. Actinobacillus pleuropneumoniae apxIA mutant vaccine
a. Vaccine Ontology ID:
VO_0002766
b. Type:
Live, attenuated vaccine
c. Status:
Research
d. Gene Engineering of apxIA
  • Type: Gene mutation
  • Description: This apxIA mutant is from Actinobacillus pleuropneumoniae (Xu et al., 2006).
  • Detailed Gene Information: Click here.
e. Immunization Route
intranasal immunization
f. Pig Response
  • Persistence: An apxIA mutant is attenuated in pigs (Xu et al., 2006).
  • Efficacy: An apxIA mutant offered a level of cross-serovar protection against A. pleuropneumoniae infection in pigs (Xu et al., 2006).
g. Mouse Response
  • Persistence: An apxIA mutant is attenuated in mice (Xu et al., 2006).
  • Efficacy: An apxIA mutant offered a level of cross-serovar protection against A. pleuropneumoniae infection in mice (Xu et al., 2006).
6. Actinobacillus pleuropneumoniae ApxIC mutant vaccine
a. Vaccine Ontology ID:
VO_0002767
b. Type:
Live, attenuated vaccine
c. Status:
Research
d. Gene Engineering of ApxIC
  • Type: Gene mutation
  • Description:
  • Detailed Gene Information: Click here.
e. Preparation
To construct an avirulent mutant strain by inactivation of ApxI toxin, the apxIC gene of A. pleuropneumoniae serovar 10 was inactivated by inserting a chloramphenicol resistance gene cassette into the downstream XhoI site of the apxIC gene for constructing the transfer plasmid. The transfer plasmid was introduced into the electrocompetent A. pleuropneumoniae serovar 10 for homologous recombination by electroporation. The mutant strain was obtained and identified by PCR and Southern blotting (Xu et al., 2007).
f. Immunization Route
intranasal immunization
g. Mouse Response
7. Actinobacillus pleuropneumoniae apxIIC and apxIVA double deletion mutant vaccine
a. Type:
Live, attenuated vaccine
b. Status:
Research
c. Gene Engineering of ApxIIC
  • Type: Gene mutation
  • Description:
  • Detailed Gene Information: Click here.
d. Gene Engineering of ApxIVA
  • Type: Gene mutation
  • Description:
  • Detailed Gene Information: Click here.
e. Preparation
Plasmid pENT1 was used to introduce the ΔapxIVA deletion into A. pleuropneumoniae single mutant ΔapxIIC via the single-step transconjugation system. The colonies with the correct PCR profile were confirmed by Southern blot and sequencing assays (Liu et al., 2007).
f. Immunization Route
Intraperitoneal injection (i.p.)
g. Mouse Response
  • Host Strain: BALB/c
  • Persistence: The LD50 data shown that the double mutant ΔapxIICΔapxIVA was attenuated by three-fold, compared with the single mutant HB04C− (Liu et al., 2007).
  • Efficacy: Two weeks after secondary immunization, all mice were challenged with 10 LD50 of homologous (WF83) (6.0 × 107) and heterologous (1.8 × 106) (S4074) virulent A. pleuropneumonaie by intraperitoneal route. There was no mouse died in groups 1 and 2, which shown a protection efficiency of 100% against homologous challenge (Liu et al., 2007).
8. Actinobacillus pleuropneumoniae apxIIIB and apxIIID double mutant vaccine
a. Type:
Live, attenuated vaccine
b. Status:
Research
c. Gene Engineering of apxIIIB
  • Type: Gene mutation
  • Description:
  • Detailed Gene Information: Click here.
d. Gene Engineering of apxIIID
  • Type: Gene mutation
  • Description:
  • Detailed Gene Information: Click here.
e. Preparation
An apxIIIBD mutant (1536∆B∆D) was selected and the correct mutation was verified by PCR using three primer pairs. The X3BD-F2 and X3BD-R2 primer pair was designed to produce products of 3720 and 4365 bps from the wild-type 1536 and 1536∆B∆D genomes, respectively (Park et al., 2009).
f. Immunization Route
intranasal immunization
g. Pig Response
  • Persistence: The LD50 of 1536∆B∆D in mice was 6.9 x 108 CFU, while that of the wild-type was 4.0 x 108 CFU, showing that the mutation of the apxIIIBD genes attenuates the lethality of A. pleuropneumoniae in mice (Park et al., 2009).
  • Efficacy: To evaluate the ability of the mutant 1536∆B∆D strain to protect mice against lethal challenge, mice were immunized with 1536∆B∆D twice and challenged with a lethal dose of wild-type A. pleuropneumoniae 1536. Eight of the nine mice immunized with 1536∆B∆D survived for more than 72 hr after lethal challenge (89% protection), while all eight of the unimmunized mice died by 72 hr after challenge (Park et al., 2009).
9. Actinobacillus pleuropneumoniae HS93Tox-/pIG-T1K vectored vaccine
a. Vaccine Ontology ID:
VO_0004607
b. Type:
Recombinant vector vaccine
c. Status:
Research
d. Host Species as Laboratory Animal Model:
Mice
e. Vector:
A mutant form of Actinobacillus pleuropneumoniae serovar strain HS93, known as HS93Tox- which lacks the genes ApxA (structural toxin) and ApxC (post-translational activating protein) (Prideaux et al., 1998).
f. Preparation
The HS93Tox- mutant of the serovar 7 strain of Actinobacillus pleuropneumoniae lacks the genes encoding the toxin ApxA and the post-translational activating protein ApxC, but still has genes required for secretion, making it a good vector. A plasmid vector was created to express the ApxA gene inside the HS93Tox- mutant of A. pleuropneumoniae, so that the bacterium acts as a vector for its own natural toxin (Prideaux et al., 1998).
g. Immunization Route
Intramuscular injection (i.m.)
h. Virulence
Because the toxin has been attenuated, this greatly decreases the potential virulence of the vaccine.
i. Description
The HS93Tox-/pIG-T1K vector strain, using an Actinobacillus pleuropneumoniae to express the AxpA gene, was shown to protect mice against Actinobacillus pleuropneumoniae challenges (Prideaux et al., 1998). ApxA gene encodes an A. pleuropneumoniae structural toxin (Prideaux et al., 1998).
j. Mouse Response
  • Vaccine Immune Response Type: VO_0001030
  • Immune Response: The inoculation with HS93Tox-/pIG-T1K induced antibodies specific to the Apx gene, and this response was boosted when the mice were re-inoculated (Prideaux et al., 1998).
  • Challenge Protocol: The mice inoculated with HS93Tox-/pIG-T1K strain expressing the toxin gene and the HS93Tox- not expressing the toxin gene were challenged with homologous wild-type serovar 7 and heterologous serovar 1 strains of A. pleuropneumoniae (Prideaux et al., 1998).
  • Efficacy: The mice vaccinated with the HS93Tox-/pIG-T1K strain were protected against both the serovar 7 and serovar 1 challenges, but the mice only inoculated with the HS93Tox- strain not expressing the gene for the AxpA toxin received no protection against the heterologous serovar 1 challenge (Prideaux et al., 1998).
10. Actinobacillus pleuropneumoniae ureC and apxIIA double mutant deletion vaccine
a. Type:
Live, attenuated vaccine
b. Status:
Research
c. Gene Engineering of apxIIA
  • Type: Gene mutation
  • Description:
  • Detailed Gene Information: Click here.
d. Gene Engineering of ureC
  • Type: Gene mutation
  • Description:
  • Detailed Gene Information: Click here.
e. Preparation
To construct the A. pleuropneumoniae serotype 2 isogenic mutant, 12 clinical isolates were tested initially with respect to their amenability to genetic manipulation via conjugation and cointegration of pBMKUΔ1 (Tonpitak et al., 2002).
f. Immunization Route
aerosol immunization
g. Pig Response
  • Persistence: In all pigs, an immune response could be detected in the detergent extract enzyme-linked immunosorbent assay (ELISA), and only in the wild-type group six of seven pigs showed elevated levels (>10 ELISA units [EU]) in the ApxIIA ELISA 3 weeks after challenge. These results showed that the A. pleuropneumoniae double-mutant strain is highly attenuated and that the group infected with the mutant strain can be discriminated from the wild-type-infected group based on the ApxIIA ELISA (Tonpitak et al., 2002).
  • Efficacy: A single aerosol application of the attenuated double mutant resulted in protection from clinical disease comparable to that obtained with two applications by using a conventional bacterin vaccine. In addition, immunized pigs were protected significantly from colonization of the lungs (Tonpitak et al., 2002).
11. HS93Tox-/pIG-T1K
a. Vaccine Ontology ID:
VO_0004637
b. Type:
Recombinant vector vaccine
c. Status:
Research
d. Host Species for Licensed Use:
Baboon
e. Gene Engineering of ApxIA
  • Type: Recombinant vector construction
  • Description: HS93Tox-/pIG-T1K, expresses the Apx structural protein in a non-activated form (Prideaux et al., 1998).
  • Detailed Gene Information: Click here.
f. Preparation
HS93Tox-/pIG-T1K, expresses the Apx structural protein in a non-activated form (Prideaux et al., 1998).
g. Immunization Route
Intramuscular injection (i.m.)
h. Mouse Response
  • Vaccination Protocol: Mice were vaccinated with alive strain of HS93Tox-/pIG-T1K (Prideaux et al., 1998).
  • Vaccine Immune Response Type: VO_0003057
  • Challenge Protocol: Mice were challenged with a wild-type serovar 7, and a serovar 1 strain (Prideaux et al., 1998).
  • Efficacy: Live vaccination of mice with HS93Tox-/pIG-T1K offered protection against homologous wild-type serovar 7 challenge, and also heterologous challenge with a serovar 1 strain (Prideaux et al., 1998).
12. PleuroStar APP
a. Tradename:
PleuroStar APP
b. Manufacturer:
Novartis Animal Health
c. Vaccine Ontology ID:
VO_0000889
d. Type:
Recombinant vector vaccine
e. Status:
Licensed
f. Host Species for Licensed Use:
Pig
g. Immunization Route
Intramuscular injection (i.m.)
h. Description
Recombinant ApxII, TbpB, CysL, OmlA(1), and OmlA(2) proteins (Van et al., 2001)
13. Porcilis APP
a. Tradename:
Porcilis APP
b. Manufacturer:
Intervet
c. Vaccine Ontology ID:
VO_0001158
d. Type:
Recombinant vector vaccine
e. Status:
Licensed
f. Host Species for Licensed Use:
Pig
g. Immunization Route
Intramuscular injection (i.m.)
h. Description
Extracted ApxI, ApxII, ApxIII, and outer membrane proteins (Chiers et al., 1998).
14. rS.C-APP-ApxI/ApxII
a. Vaccine Ontology ID:
VO_0004785
b. Type:
Recombinant vector vaccine
c. Status:
Research
d. Host Species for Licensed Use:
Baboon
e. Gene Engineering of ApxIA
  • Type: Recombinant vector construction
  • Description: S. cerevisiae expressing either ApxI or ApxII (Shin et al., 2013).
  • Detailed Gene Information: Click here.
f. Gene Engineering of ApxIIA
  • Type: Recombinant vector construction
  • Description: S. cerevisiae expressing either ApxI or ApxII (Shin et al., 2013).
  • Detailed Gene Information: Click here.
g. Preparation
The surface-displayed ApxIIA#5 expressing S. cerevisiae and the ApxIA expressing S. cerevisiae (Shin et al., 2013).
h. Immunization Route
Oral immunization
i. Pig Response
  • Vaccination Protocol: Three groups were designated as the untreated pigs (n = 5, control), the pigs fed with the vector-only S. cerevisiae (n = 5, vector control), and the experimental pigs fed with the ApxIA expressing S. cerevisiae and the surface-displayed ApxIIA#5 expressing S. cerevisiae at a time (n = 10, vaccinated group), respectively. The yeast vaccines were followed by three-time administrations with ApxIA (1.5 × 109 CFU) and ApxIIA (1.5 × 109 CFU) with one week interval (Shin et al., 2013).
  • Vaccine Immune Response Type: VO_0003057
  • Challenge Protocol: The pigs were challenged intranasally one week after final vaccination with a dose (1.5 × 109 CFU) of A. pleuropneumoniae serotype 5 Korean isolate from a pig with porcine pleuropneumonia (Shin et al., 2013).
  • Efficacy: The vaccinated pigs showed higher specific IgG- and IgA-related antibody activities than the non-treated control and vector control pigs. Additionally, the induced immune responses were found to protect pigs infected with A. pleuropneumoniae according to the analysis of clinical signs and the gross and microscopic pulmonary lesions (Shin et al., 2013).
15. rS.C-APP-ApxIIA
a. Vaccine Ontology ID:
VO_0004786
b. Type:
Recombinant vector vaccine
c. Status:
Research
d. Host Species for Licensed Use:
Baboon
e. Gene Engineering of apxIIA
  • Type: Recombinant vector construction
  • Description: A neutralizing epitope fragment of ApxIIA toxin was expressed on the cell surface of Saccharomyces cerevisiae (Kim et al., 2010).
  • Detailed Gene Information: Click here.
f. Preparation
(Kim et al., 2010) A neutralizing epitope fragment of ApxIIA toxin was expressed and immobilized on the cell surface of Saccharomyces cerevisiae for efficient vaccine development.
g. Immunization Route
Oral vaccination
h. Mouse Response
  • Vaccination Protocol: Oral immunization was preceded by overnight fasting of the mice (water was provided ad libitum). Freshly harvested 2.5 x 107 or 2.5 x 108 cells were dissolved into 1ml of 0.9% saline and orally administered at 200 µl/mouse through an oral gavage at 10 day intervals, 4 times (Kim et al., 2010).
  • Vaccine Immune Response Type: VO_0003057
  • Challenge Protocol: The immunized mice were injected intraperitoneally with 200 ml of an A. pleuropneumoniae preparation (about 1 x 108 CFU) or buffer control 10 days after the final immunization (Kim et al., 2010).
  • Efficacy: The mice fed the recombinant epitope-expressing yeast were protected from injection of a lethal dose of A. pleuropneumoniae (Kim et al., 2010).
V. References
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2. Chiang et al., 2009: Chiang CH, Huang WF, Huang LP, Lin SF, Yang WJ. Immunogenicity and protective efficacy of ApxIA and ApxIIA DNA vaccine against Actinobacillus pleuropneumoniae lethal challenge in murine model. Vaccine. 2009; 27(34); 4565-4570. [PubMed: 19520199].
3. Chiers et al., 1998: Chiers K, van Overbeke I, De Laender P, Ducatelle R, Carel S, Haesebrouck F. Effects of endobronchial challenge with Actinobacillus pleuropneumoniae serotype 9 of pigs vaccinated with inactivated vaccines containing the Apx toxins. The Veterinary quarterly. 1998; 20(2); 65-69. [PubMed: 9563163].
4. Haesebrouck et al., 2004: Haesebrouck F, Pasmans F, Chiers K, Maes D, Ducatelle R, Decostere A. Efficacy of vaccines against bacterial diseases in swine: what can we expect?. Veterinary microbiology. 2004; 100(3-4); 255-268. [PubMed: 15145504].
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6. Kim et al., 2010: Kim JM, Jung DI, Eom YJ, Park SM, Yoo HS, Jang YS, Yang MS, Kim DH. Surface-displayed expression of a neutralizing epitope of ApxIIA exotoxin in Saccharomyces cerevisiae and oral administration of it for protective immune responses against challenge by Actinobacillus pleuropneumoniae. Bioscience, biotechnology, and biochemistry. 2010; 74(7); 1362-1367. [PubMed: 20622458].
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8. Li et al., 2008: Li L, Zhou R, Li T, Kang M, Wan Y, Xu Z, Chen H. Enhanced biofilm formation and reduced virulence of Actinobacillus pleuropneumoniae luxS mutant. Microbial pathogenesis. 2008; 45(3); 192-200. [PubMed: 18585450].
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11. Prideaux et al., 1998: Prideaux CT, Pierce L, Krywult J, Hodgson AL. Protection of mice against challenge with homologous and heterologous serovars of Actinobacillus pleuropneumoniae after live vaccination. Current microbiology. 1998; 37(5); 324-332. [PubMed: 9767712].
12. Prideaux et al., 1999: Prideaux CT, Lenghaus C, Krywult J, Hodgson AL. Vaccination and protection of pigs against pleuropneumonia with a vaccine strain of Actinobacillus pleuropneumoniae produced by site-specific mutagenesis of the ApxII operon. Infection and immunity. 1999; 67(4); 1962-1966. [PubMed: 10085043].
13. Shin et al., 2007: Shin SJ, Shin SW, Kang ML, Lee DY, Yang MS, Jang YS, Yoo HS. Enhancement of protective immune responses by oral vaccination with Saccharomyces cerevisiae expressing recombinant Actinobacillus pleuropneumoniae ApxIA or ApxIIA in mice. Journal of veterinary science. 2007; 8(4); 383-392. [PubMed: 17993753].
14. Shin et al., 2013: Shin MK, Kang ML, Jung MH, Cha SB, Lee WJ, Kim JM, Kim DH, Yoo HS. Induction of protective immune responses against challenge of Actinobacillus pleuropneumoniae by oral administration with Saccharomyces cerevisiae expressing Apx toxins in pigs. Veterinary immunology and immunopathology. 2013; 151(1-2); 132-139. [PubMed: 23206402].
15. ThePigSite Pig Health: Actinobacillus Pleuropneumonia (App) [http://www.thepigsite.com/pighealth/article/309/actinobacillus-pleuropneumonia-app]
16. Tonpitak et al., 2002: Tonpitak W, Baltes N, Hennig-Pauka I, Gerlach GF. Construction of an Actinobacillus pleuropneumoniae serotype 2 prototype live negative-marker vaccine. Infection and immunity. 2002; 70(12); 7120-7125. [PubMed: 12438394].
17. Van et al., 2001: Van Overbeke I, Chiers K, Ducatelle R, Haesebrouck F. Effect of endobronchial challenge with Actinobacillus pleuropneumoniae serotype 9 of pigs vaccinated with a vaccine containing Apx toxins and transferrin-binding proteins. Journal of veterinary medicine. B, Infectious diseases and veterinary public health. 2001; 48(1); 15-20. [PubMed: 11254095].
18. Xu et al., 2006: Xu F, Chen X, Shi A, Yang B, Wang J, Li Y, Guo X, Blackall PJ, Yang H. Characterization and immunogenicity of an apxIA mutant of Actinobacillus pleuropneumoniae. Veterinary microbiology. 2006; 118(3-4); 230-239. [PubMed: 16930871].
19. Xu et al., 2007: Xu FZ, Shi AH, Chen XL, Yang B, Wang JL. [Construction and immunogenicity of an attenuated mutant of Actinobacillus pleuropneumoniae by insertional inactivation of apxIC]. Wei sheng wu xue bao = Acta microbiologica Sinica. 2007; 47(5); 923-927. [PubMed: 18062275].