VIOLIN: Vaccine Investigation and Online Information Network

Pathogen Page

Vibrio cholerae

Table of Contents

General Information
Vaccine Related Pathogen Genes
1. ctxB (Protective antigen)
2. ctxB (Protective antigen)
3. tcpA (Protective antigen)
4. TcpF (Protective antigen)
5. toxin-coregulated pili (TCP). (Protective antigen)
6. hemA (Virmugen)
Vaccine Related Host Genes
1. IgA
2. IgG Fc fragment
Vaccine Information
References

I. General Information

1. NCBI Taxonomy ID:

666

2. Disease:

Cholera

3. Introduction

Cholera is an acute intestinal infection caused by ingestion of food or water contaminated with the bacterium Vibrio cholerae. It has a short incubation period and produces an enterotoxin that causes a copious, painless, watery diarrhoea that can quickly lead to severe dehydration and death if treatment is not promptly given. Vomiting also occurs in most patients. Most persons infected with V. cholerae do not become ill, although the bacterium is present in their faeces for 7-14 days. When illness does occur, about 80-90% of episodes are of mild or moderate severity and are difficult to distinguish clinically from other types of acute diarrhoea. Less than 20% of ill persons develop typical cholera with signs of moderate or severe dehydration. Cholera remains a global threat and is one of the key indicators of social development. While the disease no longer poses a threat to countries with minimum standards of hygiene, it remains a challenge to countries where access to safe drinking water and adequate sanitation cannot be guaranteed. Almost every developing country faces cholera outbreaks or the threat of a cholera epidemic (WHO: Cholera).

4. Microbial Pathogenesis

Cholera is transmitted via the fecal–oral route through contaminated food or water. Soon after oral ingestion, V. cholerae colonizes in the small intestine and produces virulence factors, including cholera toxin (CT). Acute secretory diarrhea mediated primarily by CT causes massive dehydration and a mortality rate of 20–50% if left untreated (Xu et al., 2009).

5. Host Ranges and Animal Models

Cholera can be modeled in an infant mouse model (Kirn and Taylor, 2005).

6. Host Protective Immunity

The adapted protective immunity against cholera is thought to be essentially humoral and predominantly based on intestinal mucosal antibodies, especially secretory IgA (sIgA), which is generally considered a correlation of protection for cholera (Xu et al., 2009).

II. Vaccine Related Pathogen Genes

1. ctxB

Gene Name : ctxB
VO ID : VO_0011270
NCBI Protein GI : 110264635
Other Database IDs : CDD:279687
Taxonomy ID : 127906
Gene Strand (Orientation) : ?
Protein Name : cholera toxin B
Protein pI : 8.47
Protein Weight : 11386.27
Protein Length : 165
Protein Note : Heat-labile enterotoxin beta chain; pfam01376

Protein Sequence : Show Sequence

>ABG56901.1 cholera toxin B, partial [Vibrio cholerae O1]
MIKLKFGVFFTVLLSSAYAHGTPQNITDLCAEYHNTQIHTLNDKIFSYTESLAGKREMAIITFKNGATFQ
VEVPGSQHIDSQKKAIERMKDTLRIAYLTEAKVE

Molecule Role : Protective antigen
Molecule Role Annotation : The immune sera elicited by CTB DNA vaccine were functional and protective against V. cholerae O1 infection in mice (Xu et al., 2009).
Related Vaccine(s): V. cholerae CTB DNA Vaccine

2. ctxB

Gene Name : ctxB
Sequence Strain (Species/Organism) : Vibrio cholerae
NCBI Protein GI : AET14210
Other Database IDs : CDD:279687
Taxonomy ID : 666
Protein Name : CtxB
Protein pI : 8.48
Protein Weight : 13727.75
Protein Length : 162
Protein Note : Heat-labile enterotoxin beta chain; pfam01376

Protein Sequence : Show Sequence

>AET14210.1 CtxB [Vibrio cholerae]
MIKLKFGVFFTVLLSSAYAHGTPQNITDLCAEYHNTQIHTLNDKIFSYTESLAGKREMAIITFKNGATFQ
VEVPGSQHIDSQKKAIERMKDTLRIAYLTEAKVEKLCVWNNKTPHAIAAISMAN

Molecule Role : Protective antigen
Molecule Role Annotation : (Vishwakarma et al., 2015)

3. hemA

Gene Name : hemA
NCBI Protein GI : 12656010
Other Database IDs : CDD:178819
CDD:133452
CDD:144372
Taxonomy ID : 666
Gene Strand (Orientation) : ?
Protein Name : glutamyl-tRNA reductase
Protein Length : 419
Protein Note : glutamyl-tRNA reductase; Reviewed; PRK00045

Protein Sequence : Show Sequence

>gi|12656010|gb|AAK00701.1|AF227752_2 glutamyl-tRNA reductase [Vibrio cholerae]
MSLLAIGINHNTASVELREKVAFGPEKLSLALNQLSTSSHVKGGVILSTCNRTEIYCDVRSASKNKVIEW
LSQFHQVSLDELKPSLYVHEEQAAIRHLMRVACGLDSLVLGEPQILGQVKQAYAEARENHAVNPATEKLF
QKAFSVAKRVRTETEIGGSAVSVAYAACTLAKHIFESLADATVLLVGAGETIELVAKHLAGHHCKRMIVA
NRTRERALSLAQQFGADVIALNEIPDYLAQADIVISSTASPLPIIGKGMVESALKARRHQPMLLVDIAVP
RDIEPQVGKLNDAYLYSVDDLQSIVDSNIEQRKVEAIQAEAIVSEESATFMSWMRSLQAVDSIRDYRKQA
NEAREELLNKSLQALAAGGDPEKLLIELSNKLTNKLIHTPTRALQTAAEQGEPAKLAVIRQSLGLDDLN

Molecule Role : Virmugen
Molecule Role Annotation : A hemA mutant is highly attenuated in mice and rabbits and induces protection from challenge with wild type V. cholerae in rabbits (Ravichandran et al., 2006).
Related Vaccine(s): Vibrio cholerae hemA mutant vaccine

4. tcpA

Gene Name : tcpA
Sequence Strain (Species/Organism) : Vibrio cholerae O1
VO ID : VO_0011271
NCBI Protein GI : 21616882
3D structure: PDB ID : 1OQV
Other Database IDs : CDD:283573
Taxonomy ID : 127906
Gene Strand (Orientation) : +
Protein Name : TcpA
Protein pI : 7.25
Protein Weight : 18548.51
Protein Length : 239
Protein Note : non-toxic;
serogroup: O1; biotype: EVC

Protein Sequence : Show Sequence

>AAM66403.1 TcpA, partial [Vibrio cholerae O1]
IMGVVSAGVVTLAQRAIDSQNMTKAAQSLNSIQVAMTQTYRALGNYPATADAAAAAKLTTGLISLGKISA
DEAKNPFTGGNMNIFSFSRNAAQNKAFAITVDGLTQAQCKTLITSVGDMFPYVAIKAGAAIAFGDLTDFE
TTQADAAAGAGVIKSIAPASKNLDLTNIIHVEELCKGAAPFGVAFGNS

Molecule Role : Protective antigen
Molecule Role Annotation : Antibodies against TCP were able to protect baby mice against challenge with V. cholerae O1 of the classical but not of the El Tor biotype (Osek et al., 1994).

5. TcpF

Gene Name : TcpF
Sequence Strain (Species/Organism) : Vibrio cholerae O1 biovar El Tor str. N16961
VO ID : VO_0011272
NCBI Gene ID : 2614504
NCBI Protein GI : 15640854
Locus Tag : VC0837
Genbank Accession : AE003852
Protein Accession : NP_230485
Other Database IDs : GDB:5363132
Taxonomy ID : 243277
Chromosome No : I
Gene Starting Position : 898672
Gene Ending Position : 899688
Gene Strand (Orientation) : +
Protein Name : toxin co-regulated pilus biosynthesis protein F
Protein pI : 6.37
Protein Weight : 35844.42
Protein Length : 338
Protein Note : similar to SP:P29488 GB:L01623 PID:155281 PID:155284 PID:48417; identified by sequence similarity; putative

DNA Sequence : Show Sequence

>NC_002505.1:898672-899688 Vibrio cholerae O1 biovar El Tor str. N16961 chromosome I, complete sequence
TATGAGATATAAAAAAACCTTAATGTTATCAATCATGATTACATCATTTAACTCATTTGCATTTAATGAT
AATTATAGTTCAACCAGTACGGTTTATGCTACGTCTAATGAAGCTACGGATTCAAGGGGGAGTGAGCACC
TTAGATACCCTTATTTAGAGTGTATCAAAATCGGTATGAGTAGAGATTATCTTGAAAACTGTGTGAAAGT
ATCTTTTCCAACCTCTCAAGATATGTTTTATGACGCATACCCATCGACAGAATCTGACGGTGCTAAGACA
AGAACAAAAGAAGATTTCTCTGCTCGGCTTTTAGCTGGTGATTATGATAGTTTACAGAAATTGTATATAG
ATTTTTATCTTGCTCAAACGACTTTCGATTGGGAAATACCGACAAGGGATCAAATTGAGACACTAGTTAA
TTATGCTAATGAAGGTAAATTGTCTACGGCACTTAATCAAGAGTATATTACAGGTCGATTTCTTACAAAA
GAAAATGGTCGATATGATATTGTGAATGTTGGTGGTGTTCCAGATAATACTCCAGTTAAACTACCAGCTA
TTGTTTCTAAACGCGGACTAATGGGTACAACTTCTGTAGTTAATGCAATTCCCAATGAAATTTATCCTCA
TATCAAAGTTTATGAAGGGACTCTTAGTCGTTTAAAACCTGGTGGTGCAATGATCGCAGTATTAGAATAT
GACGTAAGTGAATTAAGTAAGCACGGGTATACCAATCTGTGGGATGTGCAGTTTAAGGTTCTTGTTGGAG
TTCCACATGCAGAAACAGGAGTTATCTATGACCCTGTTTATGAAGAAACTGTCAAACCATATCAGCCTAG
TGGCAACTTAACGGGTAAGAAGTTGTATAATGTATCTACAAATGACATGCATAATGGTTACAAGTGGTCA
AATACTATGTTCTCAAATTCTAATTATAAAACGCAAATATTATTAACTAAAGGAGATGGAAGTGGTGTGA
AACTGTATAGCAAAGCATATTCAGAGAACTTTAAATA

Protein Sequence : Show Sequence

>NP_230485.1 toxin co-regulated pilus biosynthesis protein F [Vibrio cholerae O1 biovar El Tor str. N16961]
MRYKKTLMLSIMITSFNSFAFNDNYSSTSTVYATSNEATDSRGSEHLRYPYLECIKIGMSRDYLENCVKV
SFPTSQDMFYDAYPSTESDGAKTRTKEDFSARLLAGDYDSLQKLYIDFYLAQTTFDWEIPTRDQIETLVN
YANEGKLSTALNQEYITGRFLTKENGRYDIVNVGGVPDNTPVKLPAIVSKRGLMGTTSVVNAIPNEIYPH
IKVYEGTLSRLKPGGAMIAVLEYDVSELSKHGYTNLWDVQFKVLVGVPHAETGVIYDPVYEETVKPYQPS
GNLTGKKLYNVSTNDMHNGYKWSNTMFSNSNYKTQILLTKGDGSGVKLYSKAYSENFK

Molecule Role : Protective antigen
Molecule Role Annotation : Antibodies generated against TcpF are protective against experimental V. cholerae infection in the infant mouse cholera model (Kirn and Taylor, 2005).

6. toxin-coregulated pili (TCP).

Gene Name : toxin-coregulated pili (TCP).
Sequence Strain (Species/Organism) : Vibrio cholerae
NCBI Protein GI : ORR76700
Taxonomy ID : 666
Protein Name : TCP pilus virulence regulatory protein
Protein pI : 9.2
Protein Weight : 31179.47
Protein Length : 350
Protein Note : Derived by automated computational analysis using gene prediction method: Protein Homology.

Protein Sequence : Show Sequence

>ORR76700.1 TCP pilus virulence regulatory protein [Vibrio cholerae]
MIGKKSFQTNVYRMSKFDTYIFNNLYINDYKMFWIDSGIAKLIDKNCLVSYEINSSSIILLKKNSIQRFS
LTSLSDENINVSVITISDSFIRSLKSYILGDLMIRNLYSENKDLLLWNCEHNDIAVLSEVVNGFREINYS
DEFLKVFFSGFFSKVEKKYNSIFITDDLDAMEKISCLVKSDITRNWRWADICGELRTNRMILKKELESRG
VKFRELINSIRISYSISLMKTGEFKIKQIAYQSGFASVSYFSTVFKSTMNVAPSEYLFMLTGVAEK

Molecule Role : Protective antigen
Molecule Role Annotation : (Eko et al., 2000)

III. Vaccine Related Host Genes

1. IgA

Gene Name : IgA
Sequence Strain (Species/Organism) : Oryctolagus cuniculus
NCBI Protein GI : 4063655
Other Database IDs : CDD:143182
CDD:197704
CDD:209398
Taxonomy ID : 9986
Gene Strand (Orientation) : ?
Protein Name : immunoglobulin heavy chain VDJC-alpha transrecombinant region
Protein Length : 306
Protein Note : Immunoglobulin (Ig) heavy chain (H), variable (V) domain; cd04981

Protein Sequence : Show Sequence

>gi|4063655|gb|AAC98362.1| immunoglobulin heavy chain VDJC-alpha transrecombinant region [Oryctolagus cuniculus]
METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGGSLTLTCTVSGFSLNTYPMQWVRQAPGKGLEWIGWMN
NNGNIYYASWAKSRSTITRNTNENTVTLKMTSLTTEDTATYFCARYLSSHGYDIWGPGTLVTVSSDPVNT
RPILIPLPCPILGPGEPVVIGCLIRGFFPLGPLSVTWNTSGENLTFPPVQSATSSLYTTCSLLRLPAEQC
PEENSVACHVEHNYDKGQDVTVPSPPECQPPTPGPSDTTTCPCPCPSPSCGEPSLSLQRPDLRDLLLNSN
ASLTCTLRGLKNPEGAVFTWEPTNGN

Molecule Role : Vaximmutor
Related Vaccine(s): Vibrio cholerae hemA mutant vaccine

2. IgG Fc fragment

Gene Name : IgG Fc fragment
Sequence Strain (Species/Organism) : Oryctolagus cuniculus
NCBI Gene ID : 100343867
Taxonomy ID : 9986
Chromosome No : Un
Gene Starting Position : 315920
Gene Ending Position : 352013
Gene Strand (Orientation) : +
Protein Name : Fc fragment of IgG binding protein

DNA Sequence : Show Sequence

>gi|283557656:315920-352013 Oryctolagus cuniculus breed Thorbecke inbred unplaced genomic scaffold, OryCun2.0 chrUn0214, whole genome shotgun sequence
CATGGGTTCCCTGAGAAGCTGGTGGACGCTGTGGGCCGGAGCCACCCTCCTGTGGGGTAAGTCAGACCCA
GTCCCCATCTGTGTCTTTCTCCCACTCTGCCTGGGTGGCCCATGCCCAACTTGTGCCCCTCCAGGCACTC
GCCTTCCGAGGCCTCCCCATGCCAGCTGGGATGGGAGAGAGGCCCCCAGTTCCTGCAGAGGGGCACGGCG
GGCTCTGCCCAGGCTCACTCTTCTGGAGGGAAGCCACAGAGCCCTCGGCCCCCAAAGCCGAGGGCGCAAC
AGAAACCACTGTCTTGGTTCTCTGCAACTCAGCTAAGCCATGGCTCTGAGTCTCCTTCTGCCAAGGTGAT
GGGAACCGTCACCAAGGACATTCCTGGAGAGCTCTGGTGCCCAGTTCTCCTGGGGGAACCGCCAAGAGTC
TTAGGTGAACCCGGAGGTCCCTTGACTCTTGACCCCCTGGGCCTGGGGCACACTGGGATGTCCTCCAAGT
CTCCCTGAACTTGGCAAGTTGTCACTCCAGCGCCCCCAACTCTCTCTCTATCTCTCTCAGCACTGGGCAC
TCTCTTCCTGGTAGGGGCTGTTGAGACATCCACCTTTCCCACAACCCCCTTCTTTCCTTTACCTTCTTCA
CACAAACCCTCCGGGTCGTATCAAGACAAAAGTCACCCGCCGGCCGCGGAGTGCTGCCGGTTGGGACCCC
CGAGGTGCCTCAAAACAATTTTGGGTTGAAGAGTTCACATACACATTCAGAGAGCCAGTTTCTCTTGAAC
ATGGTAAGATCTAGCACTGAAGATAAAAATAAAAATAAAAAAAGGCCGCAATCCCTCCTGTCAGCCCCCT
GCAACAGCAGGCAGCGAGCGCCCACTCTGCCTTCCACGTCGCCGCGGGACTGCGCGTCTGTGTGTCTCAC
CGGCCCCCTCAGCACCTGAGAGCCCTGTCCAAAGAGTTTGGGCTTGTAGAAAAATGAAAGCCAGAATGGC
TTGTAGGCAGGTGAGCGGTGCAGGAATAAGAACAAAGGCAAGAGAAGAGGAAAAAAAAAAAAGTTCCTGG
CACAGGGCAAAGGGTTGGCCGACTGACCAGGCGGATTAGGTAAGTGGACCAACGACTGCATTTTCTCAAG
GGTGGACCCAGGAGGCAGCTGTGGAGCCCAGGAATCCTGGGGGGCAGGAATTCCTCATGGCCTTCCTGCA
GAATTACCAGCCCGAGTACAGCGCCGCCTCCCTCTACCTGCTCGTGTCCAGCCTATCGGAGAGCCCGGCC
TTGGTCTCCGTGCTCAGCCAGGCCGACAAAGCCTCGCAGACGGTCACAGTGAAGCCAGGGGAGTCAGTCG
TGGTCAATATCAGTGCTGCGGCTGAGATGACCGGCAGCAACATCTTCCAGCACGCGGTTGTGGTCCGCTC
CGACCACCGCATCACTGTGCAGGCCCTCAGTGCCAAGCCCAGCACGGCGGAGCTGACCCTGTTGCGACCG
GTCAGCGCTCTGGGCACCGAATACTTCGTGCTCACACCCCCGGGTACCTCAGCCAACAGCCTCAGGGAGT
TTGCTGTGGTGGCCGGGGCGTCAGGTGCCTCGGTCAGTATCACCCTGAAGGGGGCAGTGACGTTCCAGGA
CAAGATCTACCCAGCAGGCGATGTGCTGAGCGTGACCCTGCAGCCCTACGAGGTGGCTCAGGTACAGGGC
GAAGTTGATCTCTCGGGATCGAAGGTCACAGCCAGTAACCCTGTGGCTGTCCTCTCGGGCCACAGCTGTG
CCCAGAAACACACAAACTGCAACCACGTGGTAGAGCAGCTGCTGCCCACGTCTGCGTGGGGCACCCAGTA
TGTGGTGCCCCCACTGGCCTCCCAGAGCCGCTATGATCTGGCCTACGTAGTGGCCAGCCAGGCCACGAAA
CTGACCTACAACCACGGAGGTACCACAGGCTCCCGTTCACTCCAGATGGGAGGCGTGGTAGAGTTTGAGG
TGCGGCCGTCCCGGCCACTCTACCTGTCCGCAGATGTGGGCATCCAGGTCCTGCTCTTCGGCACGGGGGC
TATAAAGAATGACATGACCTATGACCCCTACCTGGTGCTGATCCCGGATGTGGCAGCCTACTGCCCCACC
TATGTGGTCCAGAGCGCACCAGGCTCTATGGGTGTGGCTGTGCTGGTGGCACAGACAAAGGCTGTCAACG
GGCTGACCATGGATGGAGACACGCTGAAGGCCACGCTCCCCTGGGTGGCTGTGCCAGGCAGTGAATTCTC
ATATACTGAAGTGGACCTTGGCACAGCTAACAGGATTCACACAGTCAACGCCAACACCAACATCGGGCTG
CTCACGTTCGGGCTGGGCCAGGCTATGGGCTACGGGACAGCAGCTTCCTGCGGCCAAAGTGAGTGATGGG
AAATGGCCCCTGGCCCCCACCCACCTGCTCCTCCATGGCTTTCTGGTTCAATGGGGCTGTCCACCCTCAC
CTCCATCCTCCCCGCCATCCTTTCTTGAGTCCAGGACATGGATGCCACTCAACTTCAAGGGATCACTTTG
GGGCTTGGATAATCAAAGAGAGGAGGAATCGATGATCCATCTCCGTAGCTAACACTCACTGTGACACCTG
GGTTCACTTCCAAATCTTACTTCTGTTTTAGCTTCTTCAGGGGCGGGCATTGTGGTGCAGTGTGCTGAGC
TGCAGCTTGGAATGCCCACGTCCCAAATCGGAGTGCCAGTCTGAGTCCCCCCACCTCTGCTTCTACCCAG
CTTCCTGCCAATGCATCCAGTGGGGGGAGGGGGAGGCAGCAGATGGTGGCTCAAGTACTTGGGTCCCTGA
TGCCTATTTGGGAAACCCTGTTGAAGTTCTGGGCCCCTGGCTTAAGACTGGTCCTGTGCTGGCTGTTACA
GGCACTTGGAGATTAAGCCAGCGGATGGAGTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTC
TCTCTCTCTCTCTCCCTCTCCCTCTCCCTCTCCCTCTCTCCATCTCCCCCACTCACTCTCCTCTCTCCCT
CTCTCCCTCCTCCTCTGCCTTTTAAGTAGATGAAAACAACTAAACATCTTTTAAAAATAAGCAAAACCAT
CTCCACACCTCTGCCAGATTCCCCTATGGAGATGGGATTGCTATCCTATTCCCCTCATCCTACGAATGAG
GCAACTGGGGCACTAAGAGTTTAAAAATAAGTTGCCCGGTGTCTCCGAGCTGCATGATGCAGCCCGGATT
CCTAGCTAGACAACGTGACTCCACAGTCATTGCCACTGTGCTGAGCGGCCTCTGCCTTCTACTTTCTGCT
CTTCAAAAGATCCCTCAGCCTTTAGAGCCTGGTGGGATGGCAGGGGTGAGGGGGAAAAGGGAGGGAATAC
AATGAAGATGTCAATAACAGTTCCTGTCACTCCATCTGCACTGCCTTCCCTTCTGCTCAGCTGCTCTGTG
TTAAACAGGGGGATCAGAAAGCCCAGGCCTGAGCAGCTCTGTCTCTGTGTGATGCCAGCCCTCAGTCTCC
CCATCTCTAAAATGGGCAGCTATGAATGCACTCCTTGTGGTGAGAGAGACCCAGGACTCCTGCCCCTGTG
GGCATTTCTTTTTATTTTTTCTGAGGCAGTGAGAGAGAGAGATTTCATCAGTTGGTTCATTCCCTTTGCA
AAGGGCCAACACAGTGGGAGCCAGGAGGCAGACCTCAATGCAGGCCTCCCACATGGGTAGGGGAACCCAG
CCCCACTGCCTGCCAGGGTCTCCAGCCAGCAGGAAGCTGTAGCCAGGAGCCAGAGGTGGGAACTGGCCCC
AGGCACTCTGATGTGGCACGTAGGTTTCTGGACCATTGGGCTAATGGCCCCCGCTGGCCCTGGGTGTGGC
TGACTCTGCTCCCACCCTCTTCCCCCCAGGCCTGGTGTCCTCGGAGGAGCCCTCCTGTGAAGGCGTGACG
TGCGCGGCAGGGCAGCAATGCCAGGTGGTGGACGGGAAGGCCGTGTGCGTGGCAGAGTCCACGGCCGTGT
GCCGCGCCCAGGGCGACCCCCACTACACCACCTTCGACGGCCGGCGCTATGACATGATGGGCACTTGCTC
CTACACGATGGCCGAGCTGTGTGGCTTGGACGAAACCCTGCCTGCCTTCAGCGTGGAGGCCAAGAACGAG
CACCGGGGCAGCCTCCTGGTCTCCTACGTGGGCTTGGTCACCGTGCGTGCCTACAACCACGACGTGACAC
TGGCCCGCGGGGAGGTCGGCTTTGTTCGGGTGAGTGTCCGGGGGCTTGGGGGCTTGGCCCCCTGGGAGCC
GTGAGTCTGAAGGCAGAGGCAAAGGCCTTTCTGTGGGCTGGGGAGTGGGAGACGGGGAGCGGGGGTTAGA
AAACAATAGCTGAGCCTTAAAACCTTTCAATGCCACGTCTAGAAATTCCAAAGATGCTCAGAAAGACAAT
TAGAAAAACAAATATGCCAACATGGTGTACAAATGAGGAATTTCTTTCTTTCTTTCTTTCTTTTTTTTTT
GCTTTAAAGATTTATTTTATTAATTTTAAAGAGAGTTACAGAGAGAGGTAGACAGAGAGAAGAGAAAGGT
CTTCCATCCGCTGGTTCACTCCCCAGGTGGCCACAACAGATCGGAGCTGTGTTGATCCAAAGCCAGGAGC
CAGGAGTCTCCTCCAGGTCTCCCACATGGGCGCAGGGGCCCAAAGACTTGGGCCATCTTCTACTGCTATC
CCAGGCCATAGCAGAGAGCTGGATCAGAAGAGGAGCAGCTGGTACTAGAACTGGCGCCCATATGGGATGC
CAGCACTTCAGGCCAGGGTGTTAACCCACTGCGCCACAGCGCTGGCCCCATGAATGAGGAATTTCTTGGG
ACCAAATCACTGACCTATTTAAGTGCCCCAGTTGGTTCTTAGCTCTTAATGCTCACAGCAACTCTGGGAG
TAGGCCTTGTTATTACGGACATGCACAGAGGAGGGTACAGGGTGGGAAAGGTGAGTGATGCCCCAAGCAC
TGGGAGCTGTAAGAGCTGGCGCTGGGAACGCAGGCAGACTGGCTCCCAAGTCCCCGTTCCTCGTCCCGAG
TCTCTCTCTTCCACTGCTTGTCCGTGCTGTGCCGTTCCGCCAGTACCAGCAGCATCTGGAGCAATGACTT
CCTTTTTTTTTTTTTTTTTTATTTTGACAGGCAGAGTGGACAGTGAGAGAGAGAGACAGAGAGAAAGGTC
TTCCTTTGCCGTTGGTTCACCCTCCAATGGCCGCCGCGGCCGGCGCGCTGCGGCCGGCGCACCGCGCTGA
TCCGATGGCAGGAGCCAGGAGCCAGGTGCTTTTCCTGGTCTCCCATGGGGTGCAGGGCCCAAGCACCTGG
GCCATCCTCCACTGCACTCCCTGGCCACAGCAGAGAGCTGGCCTGGAAGAGGGGCAACCGGGACAGAATC
CGGCGCCCCGACCGGGACTAGAACCCGGTGTGCCGGCGCCGCTAGGCGGAGGATTAGCCTAGTGAGCCGC
GGCGCTGGCCTGATTTCCTTTTTTGTCACGTTATTATGAAAATGTTAGAGCATCCAAAGCAGGTGAAATC
ACCAGACAGTGAACCTCGTGGGCCCAGCGCCTCAGTTCCAAGCTTTTTGCTATTCTTGGCTGATTAGACC
TCTGTCCACACAGTCTGCCCTCCCTCCAACAGCCGCTCTTGTCTATGTGCCTGGAAGTAAGCTCTGCACA
CCAGCTCCACTTTCTGTAAGGTCTCCTCATGCAGATCACCCAGAAGGGTGCACTATTGTAAAACTCCCCC
GTTTTGTTAATTTTAAGATTTTTAAGAAAACATTTAGTTTGTAACAGATTCAATATAGCTTTTAGATATG
ATTCTAAGGATATAATGACATTCCCTTCCTCCTTTTTCCATGTTTTCAAGATTAAACTTACATATCACAA
AATTCACAAATTGCAAATGAGCAGTTTGCTGTTTTGGCAAATGTGTAACCCAGAGTCCCATTAAGATACA
GAATATTCCCATCAACTCAGAAAGTTCCCTCCCGAACAACCCCCAGCCCACTGCACAAGGCACAGTTCTA
ATTTTTTTTCCCCACCATGATCTACCGTGGATCAGTTTTGCTTGCTTAAGAATGTCCAATAAAAGCAGGC
ACACAGCTTAGCAGGCAAAGAATTCACATAGCACAAAATAAAAGGGGTAAGATACAAAAGATCTTGTTTC
TATTCCAGTCTTCATCTACTCCCCTCCTCCGCCCTCCCCCCCACCCTCCCCCCACTCCTTACAGATAACT
TCTCCTATTGGTTTCTGGAGAATTCATCTAGAGTTTCTTCCTGCCAATAGAAGCAAATACAAGCGGATGA
CTTACCTTTCCCCTTTCTTAACAAAATGTACCCTACATGCTCCATTCTGCACTTGACCGTACCTTCTGGG
CCCCTTTCCCTATCAGCACGTAGGGAGCCTCCTCTCTTCTTTTGGGTGCGGGGATGGGGGAGCTGCACAG
TTCTGCATGGGATGAATGTCCCAGAGTTCACTGAAGCAGCGTGGGAGGGATGGATACTGTTCCATCTTGC
AAACAGTGCTACAATGTATATTCTTACACCTGCTCAGCCTGTGAGTGTGCAGGTGGATCTGGGAGACCAA
CAACCGGGTGTGTTACAGCAGGTGTTCTAGCGGGATAGGGCACAAGCTCAGGGGACTCCCGAGCCGCTGC
AGTTGTGTGCAGCTTCTCATGGATCTTCTGTCAGCTCCTCACAGAGGTCACTGACTTTCTCAAGAGTTGG
CGCTCCTGGTAGAATCGTGGTAGGATGTGCACAAAACATCGTGAGAAGGAGAGAGAGATCATGAAACTGT
GACTTCTGCATGGTGGCATTATCTTTTTTCATTAAAGTATAAAAGCATTGAAAAGTAGTTGGAATAACAA
NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNNCCCCTGGACTGAGAAGCTGTGAGCAGGGGTCCCTAGCACTGGCTGGCGTGTTAC
ATGGATGCATAGTAGGACTCCTGGTGGCTTTGCTGCAGGCCTGGGTGCGTGCATTCTGTCTTCTCCTCCT
GCCATTGCTGAGGCTGTCCCCTCTGCCTGCTGCTCCTGTTTTTGGAGCTCTGGCACTTGGCTAAGACGTT
CTCATCCCTCAAGTCTCAGCTCTAACATAGCCCCGGACATGTGTGAGGAGTGAGAAAGAAGTGTCTGCGG
ACACACCGAGATGCTCCTGGCTCCTGGTATCTCAGGCAGCCCTGGCTGCCACAGCAGGGGTGGCTTCTCA
ACAGCACAAGCTTCTAGCTCTGCTCTACAAGCTGGGAGCCTGAGGTCAGGGTGCCGGCATGGTTGGGTTC
TGGTGAGGGCCTGCCTGCAGCCTGCAGACTGCAGGGTTCTCACTGTGTCCTGACCAGAGAGCGCTCTGCG
GTGACATCATTCACCCATTGTGCACCTGCTGTTTTTACCAGACACAGTTTCTGCCTCTGTGGATAAAGCA
GCAAAGTGCCTGGGTAGCGCAGATGTCTGGTGGACTCAGACTGACAATAAACAGGATGAATTTGCAAGTT
ACTTTTTAGAAGGCAAGAAGCTATTCTTAACAAATGAAACATGTCAGGGGAAAGACATAGGGAATGGAGA
GGGCTCGCCATGTGAAACAGGGCTGGGATGGGCATTTAGCCTGGCAGTCAGGATGCTGGTTCAGACGCTT
GCAGCCCACACTGGAGTGCCTGGGTTCAAGTCCCAGCTTGGGATCCTGCTCACAGCTTTCTGTTCATGAA
GACCCTGGGAGGCAACGTGGGTCCCTGCCACCCATGAGAGAGACCTGGATTGAGTTCCTGGCTTCCAGCT
TTGCCCCAGCCCAGCCCCAGCTGTTGCAGACAGTTGGAGAGTAAACCAACAGATGGGAGTTCTCTCTCTT
TCTCTCAAATAAATAAAAATGTAAATAATTAAAATAATAAAAAGAGAGAGTGGCCTCACAAATATGGGAA
CAGCTGAGCAAGTGTTTTTTATTTATTTGAAAATAAATAGGAGAGAGAGAGAGGGAGAGAGAGATAGAAA
GAGATCTTCCATCTGCTGATTCATTCCCCAGATGTCCACGACAGCTGGAGCTGGGCCAGGCAGAAGCCAG
GAGCACCTTGAGGCAGGAAGCCCAGGGACATCCTCCACTGCTTTCCCAGACACATGAGCAGGGAGCTGGA
TTAGAAGTGGAGTAGCCACGACTTGAACCCGTGCTCGTATGGGATGCCAGTGTTGCAGGCGGCAGCTTAG
CCCCTTATGCCACAACACCAGCCCTCCCTGAGCAGATATTTTCTGTGAGCCAAGGAAGTGAGCCACATGC
TCATCTTGGGCAAGATCACTCCTGGCCACAGGAAGTGGGAGAGCAGAACTGTGGAGGCACAGAGGGTGGC
AGTAACAAGGAGGGATCCCAGGACAGTGAGACCCGGGCCAGGGAGCAGCCAGCCCACCCTGGGCTCTGAC
CACCCCACCTCACCCCACCTGCAGATCAACAACCAGCGTTCAGGCCTGCCAGTCTCCCTGAGCAAGGACC
GCCTGCGCGTGTATCAGCGCGGGACCCGGGCCGTGGTGGAGCTGGACTTCGGGCTGGTGGTCACCTACGA
CTGGGACGCTCAGCTGACCGTGAGCCTGCCTGCCAAATTCCAAGACCAGGTGTGCGGGCTGTGTGGCAAC
TACAATGGCAACCCCGCCGACGACTTCCTCACACCCGACTGGGTACAGGCCTCGGATGCCCTGGAGTTTG
CCAACAGCTGGAAACTGGACGATGGGGATCCCCTGTGTGATGATGGTTGCCAGGGCAACTGTCCCTCCTG
CACCGCAGCCCAGACCGAGCAGTACAAGGGCGACCATCTGTGCGGCATGCTGACCCTGGCTGAGGGCCCC
TTTGCCCCCTGCCATGAGGTCCTGGAACCCCAGTCCTTCCTAGATGATTGTGTGTACGACCTGTGTGTGA
CGGGCGGGGAGCGGCTCAGCCTGTGCCGCCTCCTCAGCACGTACGCCCAGGCCTGCCAGGAGCTCGGTGT
CTCTGTCACGGACTGGAGATCGTCAGCCAGCTGCCGTGAGTGATGCCACTAGGGGCGCCCGGGGCACATG
GGGATAGACAGGCGGGCCTCTTGGGTCTGCTTAGAATCTGAGCCCTTAGTTTGCTTGAGACTAATGCCGA
GCCTATCTGGGGCACAAGGATGGCCAAGACAGCCAGGGTCACGGCAGGCATAGCAGTATACGGCGACACC
CTGGAGCCGTCAGGCTGGGGGTGGCAGAGAGGTTGAGGCTGGGTTGGGGAAGTTCAATGGCATTGCAGGA
GACACACAGTTCTTGATCTAGATGAGGGGTGTCAGGAAGTGTTTCCTAAAAAAAGGGGTGCCCAAGTTCA
AAGCCACAGGAAGAAGTAGGAAAAAAGGAGGAGAAACTGGGTTCCTGGGGGACAGCACGTAGAGGCTTGG
CAGGAGAGGAGATCTGCAGGATCCCAACAGAGAATTGTGCAGCCTCTTCCCACTCACCACCCTTGTGTGA
CTCGCTTCTCCTCCCTGGATCAGGTCCAAGCCTTTTGGCCAGGTGTCATGCACTTCAGTCACTCACTCAT
TCAGTAAATGAGCCTTCCTTGGGCCCCCTGCCTGGCCCTGCATCACCAGTGGGTTCGTCCATCTCCCTTC
CCCCTTTCATTTCCACATCTGCCTCATTTCTGAGCACCTGCTGGGTGCCGGGCCCCTGTCCATGGTGCTG
AAGACTCAGCAGTGACTGAGGCAGACCAGGTCCCCACTGCCACAAAACTTACGTGCCAGGGAGAGAAGGG
GGAGACAGACAGCACGCAGGTAAAGTGTGAAATAGCTTCAGAGAGCAATAGGAGGAAAGTTTGCACGGTG
GAAGTGGCTGCGAGACTCCTAGAGCAGGGATGGCCTTTGTGAGCAGGTGGCATTGGAGCTGAGATGGGGC
GGGGGGCAGGCAAGGCAGGCAGCAGTGATGTTTGTGGAGCACTTCCACATCAATAACCCATTTAATCCTC
ACAACAGCTCTGTGAGGCCGGTGCTATAGCATCCTATTTACCAACAGGGAAACCAGACAAGGGGAGATGG
GAGTCATTTGCTCACCGTCACACAGCTAAGAAGGAGCAGAGCTGAGATCTGAACCCAGGCTGTCTGGGCT
CCAGTTGGCATGCTCTGTGGATGCCGTGCTGAGGGAAGGGTACACCAGGCAGAGGGAACAGCCAGTGCAA
AGGTCCTGAGGACGGAGGCCACTGTGACTGGGGCAGAGAGAGAGGGAGAGGAGATGAGAGCAAGGGAGAG
GAGATGAGAGCAAGGGAGAGGAGATGAGAGCAAGGGAGAGGAGATGAGAGCAAGGGAGTGGTGGAACAGA
TGGTGCAGGGCCTTGGGGTGACTGATGGAACCTCAGGAGGGCCCTAAGAGGGGACAAACCTGATCTGATT
CAGGTGTCCCTGGCTGCTCAGGGCAGGCAGGTGGGAGTCCAGGCAGCAGGCAACATGCTGGATAGGTTTT
GAATATGGAGTTCCCAGGACACACTGGCAGGTTGGTTGTCCAGTGGATGGCAGTGGAGTGGGAAGGGAGC
TGCTCAAGGCAGAGCTGGGGGTTCGGGCCTGGGCCGTTGTCATTGACTGAGAGGATAGTGCTGAGTTTGC
AGCCAGCAGAGGGGGGTTGCTGGGGGAGCCTGGACACTTGGCCTGACCCACCCTCATGTCTCCCCTGCAG
CCTTGTCCTGCCCCGCCAACAGCAGCTATGAGCTCTGTGGCCCCGCCTGCCCCGCCACCTGCAACTCCGT
CGCAGCACCGTCTAACTGCTCTGAGCGCCCGTGCGTGGAGGGCTGCGTGTGCCTTTCGGGTTTCGTGGCC
AGCGGCGGCGCCTGCATCCCTGAGTCATCGTGCGGGTGCACCTTCGAGGGCCTCCTACTGGCACCTGGCC
AGCAGGTATGGGCTGACACCCTGTGCAAACAGAGCTGCACCTGCGATGGCACCACCCACCAGGTGAGCTG
CCAAGACACGCAGGGCTGCCCGGAAGGCGAGCGCTGCCGCGTGGAGAACAACCTCCTGGGATGCTACCCT
GAAAGCTTTGGTAGCTGCCAGGCCTCTGGGGACCCGCACTATGTGAGCTTTGATGGCCGGCGCTTCGACT
TCATGGGCACCTGCACCTACCTGCTGGCGGGCTCCTGTGGCCAGAACACAGCACTGCCCAGCTTCAGCGT
GCTGGTGGAGAATGAGCACCGGGGCAGCCAGACCGTGAGCTACACAAGTGCCGTACAGCTGAAGGTCTAT
GGGGTGGAAGTGGCCGTGAACCGGAAGTACCCTGGGCAAGTGCTGGTGAGTGACATGGGCCCAGAGCTGG
GCAGAGGGGAGGGAGACCCCCGGGATGGGAGTTGGGGCCTGCTCCTCCTGGCCTGTGTGCCTTAGTCTTT
CACTGCCTGAGTTTTGTCCCCTGTTGAGAAGCATGAGTGGGGGCAGTTCCCGGGCTGGTTGTGGGGACTT
GGGGAGTTGGTCGTTGAGTGGTCTTTGAAAGAAGCCTGGGGGCCTGGCAGGAGCTCAGCAAATGTTGGCT
ACTGCTTATTGTTGTTGTTGTTATGCACACTCGGAACTGTAGACCCGTGCGTTTTCTGTAGGATGCTGGT
TTTTCAGGGTGCAGGTTGACATGTCGGGTGACAGGTGAGATGCCGGGAGCTGGAGGGGAGTGGATCACAG
AGGTCCTTACCCGATGTCTCCAGGGATGCTGGTGGGGAAGGCCTCCCAGGGGCCCCAATGGTCCCGGGTT
TCTGTCCAGAGGATCCTAGAGGGACTACTGCCCAGTGGGAGCCAAGGGCTACTGGCTTTGTGTTAGAAGG
TAGACATGGAGACGTGGGACTGTTGTGGCTGGTATTTGGTACTGAGCAGTGGCCTGTAGCCTGGGGAGAG
GGGTGGATGATGTGTCCAACACCACTCTAAACCCTTCATACATCTTAACTCATCACCCTCACGACAACCT
CGGAATAAAGGAAATGCTGCCCCCAGAGAACACTCTAGAGAAACGGGACAGAATTTTGACTGTATTTCTA
GTCACTGCTCTGAGATGACACTTGGGCTGAGGCTTGGAGAATACCAACCAACACATACTGGGTTAATTTA
ATGTGCTGTATCCATATGCATCAACCTGTTTAATCTGTACCACAACCCTGTGAAGTAGCTGGTCTTCAGT
GAGACTGGGCGGGGCTTCAGTGAGGATGGGCGGAGTTCGGCGAGACTGGGCGGGGATTCAGTGAGGCTGG
GCGGAGTTCAGTGAGGCTGGGCGGAGTTCAGTGAGGCTGGGCAGGGCTTCAGTGAGGCTGGGTGGGGCTT
TGGTAGAGTTGGGTGGGGCTTCGTGTGCTCAAGATGCCCGCCTGCCTGGAGTGGCAGGAGTTGGGGGTGG
GCCATACTGGGGGGTTAAACTGGGGTTCTTTCAGGCCATTGTAAAGGGTTTGGATCAAAGCTGAACCTAA
CAGGTGAGGTGGATGGAAGTGGTCCCGGGAAGAGCGATGAGGGCATAGCAAGTGCAGAGGTCCTGAGGTT
GGGGATGGGAATGCATTTGATGTGCCCAGAGACACAACAAGAGGCCAGAGCGGCTGGGGCTGAGTGAGTG
GTGTGGACACAGGAGTGGACAGGGCAGGGGTCAATCAGGACTTTATAGGGAAGGGGCCTGGATTGAAGGC
AAATCCCAAAGGCGGGGAGGAGGGGGTAAGTGTGATGATCCTGAGGCCAGGCTGAGCTGGGGCAGCTGAG
GGCTCCTGAGCAAGCCCACAGGGCTGGAGCAGAGGGAGGGGCAGAGTGGGAGGAGGGAAGCTCAGGGGGT
GGCAGGGAGACAGAGCAGGCAGGCTTTGAGGGCCCTGAGTCAGAGGTTTGCATTTAAGCTGATGCCTGAA
GAACCAGGTCAGTGGACGGCGTTCCAGGCAGAAGGAAACAGCAGGTGCCAAGGCCCTGAGGCAGAACTGA
GCATGTTACCTTCCAGAGAGTAGCAAGGAGGCCAGGGAAGCTGGAGCCAAGTGGGAGGGGGGTGAGCCAT
GGGGCCAGGAATGCCTGGCCTCTGCACCCTCCTGCCTCTCCCCCCAGGTGGATGGCGTCCTGAAGAACCT
GCCCTTCCAAGCAGCCGATGGGCGGGTGCAGCTGTTCCGCCAGGGCAACAATGCTGTCGTGCGCACGGAC
TTTGGCCTGACTGTCACCTACGACTGGGACGCCCTGGTGACCGTCAAGGTGCCCAGCAGCTACGCGAAGG
CCCTGTGTGGACTCTGTGGGAACTTCAACGGGGACCCCAGCGATGACCTGGCTCTGCGCGATGGGAACCA
GGCTGCCAATGCTCTGGACTTCGGGAACAGCTGGCAGGAGGGGACGAGACCTGGCTGTGGGGCGAGTGAC
GCCGGGGACTGTCCCACGCTGGACTCGCAGGTGACCCAGCAGCTGCAGAGCAAGAAGGAGTGTGGGATCC
TTGCTGACCCTGAAGGACCCTTCCGGGACTGTCACAAGAAGCTGGACCCCCAGGGTGCCTTGCGCGACTG
TGTGTATGACCTCTGCCTGCTGCCTGGCCAGTCTGAGCTACTGTGTGATGCTTTGGCTACATACGCTGCT
GAATGCCAGGCTGCCGGGGCCACCGTGGAGCCCTGGAGGAGCGAGGAGCTTTGCCGTGAGTACCAGAGTG
GCGAGGATGTATGTATGCGTGAGCATGTCAATATCATTCATTCACTGAATCTTTAGAACAGCGCTGTGAT
GCAGGGGCTGTGGTTCTACTCACTTAGGAGATGAGGAAAGTGAGGTTCAGCAAGGCAGGGCTGCAGGGTC
ACCACAACCAGCAAGTCACAGATGGTAGGCGTGGCCGCCAATGTTTGTGAAATGACAGGTGACATCCAGC
GGCCGCACGTCTGAAGCCCACGTGGCAGCTGTCACTCAGAGGGCATCAGCGGTGCTGCCTTAGAAGAGGC
ACGGCCCCCATCCAGGTCCTCCTGGTCCCCAGCTGGCCTGCTCCACTCACCCCACCTGCCAGGCCCCTTG
GGCAGCGCGTCTGAGAGCCTGGCTGTGGATGCGGGTGCAGGGATGACCTGCAAACGTTTTTGCTCACCTG
TGCATTTGGAAAACCACGCTTCCCCTCACCTGATTTTTTTAAAAGTTTATTTGAGAGGTAGAGTTACAGA
CTATGAGAAGGAGAGACAGAGAGAGAGGTCTTCCATCCATTGGTTCACTCCCCAAACGGCCACAACAGCT
GGAGCTGAGCCCATCCGAAGCCAGGAGCCAGGATCCTTTTCTGTGTCTCCCACACAGGCGCAGGGGTCCA
AACACTTGGGCCATCTTCCACTGCTTTCCCAGGCATAGCAGAGAGCTAGATTGGAAGAGGAGCAGCCAGG
ACCAGAATCGGTGCCCATATGGGATGCCGGTGCCTCTGGCAGAGGATTAGCCTACTATGCCACAGTTCAG
GCCTCACCCTCACCTGATTTTTAAAAATCTTGGGATACAATGTACATATTCAAAAAGACCTGGGTAAGGG
AACCACTTGATCAGTTTTTCAGTAAGAGAGCGTATCTGAGTAGACAGCCCACGTTCAGAGAAAGAACAGG
GGCGGGCGTTTGGCACAGTGGTTAAGATGTCACTTGGGATTCCTGTATCCCATATCATAGTGCCTGGGTT
CAAGTCCTGACTCTGCTTCCGACACACCTTCCTGCTAATGTGCACCCTGGGAGGCAGCAGTGACAGCTCA
AGTGCTTAGGTCGCTGCCAACCACATGAGAAACCCTGATGGAGTTCCTGGTTCCTGGTTTTGACCTGGTC
CATCCCTGGCTGTTTCAGGATTTGGAGAGTGAGCCAGCAGATGGAAGATGTCTCTGTCTCTTTGTCTCAC
TGTCTCTCAAATAAAAATGAAAATAAATAAATGTACAAAAATTTCAAATGAGCATACCAAGACCCCCATC
CCAAGAGGTAACCAATATTTTGACTTCTAAAATTGTTGAACTTCATATAAATGGAAACATACATCATTAG
CTTTCTTCTTTGGTGTAAATAATCACAAAAGATATAATTCCCAACAAATAGTAACTTTTTACTTTTAAAA
TACATTTTATTATGGAAAAATTCCAACATACCCAAAAGTAGACCAGGATGATGAACCTGCATATATCCAC
CAGAAAATTGCTACCATTATTAATATAATGTTGACATCTTAGCAGGCAACTGTTACTGATGTAACTCTCT
TCAATGTATTCAACAGAGACACAGTACACAGTAGAGCCCACCACTGCCCCCTGCAGATTCCATGAGAAAT
GGGTGGATGCCTTCAACCGTCCATCCACCCGTCCGTTCACTGATCAAATGCACTCGTGTACCATATAATG
GTGTCTCAGTCAGCAGTGGGCTGCATATACGATAGTGTTTCTGTGAGACTGGCGGGGCCTTCACTGTGTT
AGTTTGTGTTAGGGCACTTCATGGTGTTCACACAACTAAACTGCTCAATAATGCACCTCACAGAACATGT
GCCTGTCATAATGTATTTGTTGGCCCTGTGCCACATCCCAGGCAGTAGGGGTATGACAGTCCTTTTTTAA
AAGATTTATTAGAAAGAGTACAGAGAGAGAGAGAAAGAGACACTGGTTCACTCCCCAAACAGCTGCAACA
GTCAGGTCTCGTCTAGGCCAAAGCCAGGAACCAGAAACTCCATCCAGATCTCCCATGTGAGTAGCAGAGG
CCCAAACACTTGGGCCATGCTCTGCTCTTTCCCAGGCACCCAGCAGGGAGCTGGATCCGCAATGGGACAG
CCAGGATTTGCACTGGCGCTCCAATATGGGGTCCGGTGTCGCAGATGGTGGCTTAACACCCTGCACCACA
GTGCTGGCCCAGCAGTGAAAGCCTCTGTCTTCCTAAACAATGATTATTTGTTCTAAAAGAATTTTTAAGT
GATTTTTTACCATTTGCTTATTCTTTATTTGAGGGAGGGAGGGAGGGAGGGAGAGAGAGAGAGAGAGATC
TCCACTGATTTACTCCTCAAATGCCTGCAAAAGCCAGGGCTGGGACAGGCTAAAAGCTAGAAGCCTGGAA
CCCGGTCCAGGTCTCCCACATATGTGGTAGGAACCCAACCGCTTTAGCCATCACCACTGCCTCCTAAGGT
GCAAAGCTAGGATTTGAACCCGGGCACTCCCATGCGGGGTGCAGGCGTCCAAAACAGGAGCTTCATCATG
GGACCAAGTGCCTAACCTAAAAGAGATCTTTCTTTTTTAAAAATGATTTATTTATTTGAAAGAGAGTTAC
AGAGAGAGAAGGAGAGACAGAGAGATCTTCCTTCTGCTGATCCACTCCTCAAATGGCTACAACAGCCAGA
GCTGCACCAGGCCAAAGCCAGGAGCTAGGAGCCAGGAGCCAGGAGCCAGGAGCTTCTTTGAGGTCTCCCG
TGTGGGTAGCAGAGACCCAAGCACTTGGGTCATCCTCCGCTGTTTTCCAGGCACATTATCAGGGAGTGGG
ATCTGAAGTGGAGTGACGGGGACTCAAACCGGTGCCCATATGGGATGCCAGTGTCAAAGGAGGTGGCTTA
ACCCACTGCGCCACCTGCTGGCCCCAAAGATATCTTTCCATGAGACGAATGGGACTCTGCTTCACCAGTG
TTTCACACACATTGTTAAAATGAGGCCGGGGCCCCCATCCAGGCTGTCCCAGGGCGCACTTTCGCAGGAA
CAGGATTTTAAGCGGGTGGGGGTGGCCGGAATCCTGGCGAGACCTCTGTCCACTAGTTGAATCGCTTCTG
AGTCACAGGGAAGGAGCACAGGACACTAGAAAATCCATGGGTGTTTTTGGTGTGCATGTGATTTTTGAGC
GGCCGATTCTATCAGGCCCTTATTCCATCTTTTTGGGAGCAGAGAACTGGAAGGAATTTTTTGCCCATAA
TGATGCATCACCCTTTGTCCTCACAACCTTAGTGACGTTTGCAACCATGTCCCTTTCTGTCCCTGCGAGT
TCTCTCACCCCAGGGCGGCACTCCGGGGTGAGATTTGGGAGGAGAGGAGGCTGCACTGTTCTCCCCTGTC
AAGTGGGAGCACGTGCTTGGGAAAGGACACTGGGATGCTGAGAAGCGGGGATGCCCCCTGTGCCGGCACA
TCTCAGGCCGCCATTTAAGGAAGGGCCCGCATGGCGCTAGCTGGCGCTGGTGTAACCAGGCTCTGTTCTG
GGGGCTTCATGTCGTCTTTGACTCCTCACTGTCACCCTGTGGGGCTGACTCTGTCCTGTTTCCATTGTAT
AAGATGAAGAAGTGCCCAGAGATGGTGAGTAACTTGCCTGAGGTCACCCAGCATAGCACAGAGACTTGAG
CCCAGCAAGTGTGTCCTGTGTCCTGTGAATTGCCACACTCGGTGATGCCTGTGCAAATGGACACTCTCTT
ATTTCCTTTAACAGCTCTTGGCCCTTGCACCTTCAGAAGTTCTAGAAGGAGCAGGGCAGGGCCTGACCCT
ACGCCCCCCAGCCTGAGCTTCTGTGGTCATTAGAACTATCTTCTGCCTCCTGTCACTCTTCATTTTTTCT
GAAATATTCTAAAGCAAATTCTAGACATATGAGCTTTTCTATAAATACTTCAGCAAGTGCTTGCTATTCA
GTGACAGAAAGGGGCTTTTTTTTTTTTTTTAAAGAAACTTCATTTATTTGAAAGGCAGAGTTACAGAGAG
GTAGAGGCAGAGAGAGAGGTCTTCCATCTGCTGGTTCACTCATCAGATGGCCACAATGGCCAGAGCTGCG
CCGATCCAAAGCTAGGATCCAGGAGCTTCTTCCGGGTCTCCCACATGGGTGCAGGGGCTTAAGGACTTGG
GCCATCTTCCACTGCCTTCCCGGGCCACAGCAGAGAGCTGGACTAGAAGAGGAGCAACCGGGACTAGAAC
CCGGCGCCCATATGGGATGCCGGTGCTGCAGGTGGAGGATTAACCTAGTGTGCCACTGTGCTGGCCCCTT
TTAAGTCATTTTTATTCTGTAAAAACTTCCTTCCTTTCCCCCCAACACATGTGGGACTGCAAAAAAAGTT
CATGGGAAATATGTATTATGAAAATAAAAACTGCACTCAAATAAGCTTTTCTTTTAATTCCACCTTCCAC
GACCCTTCTGAAATGCCCTTATATCATTATGAAGTTTTTTAGATGTAAAGAAAAGTTGAAAGAATTTTAC
AGTGATTGTCAATACCCCCCCAGCCCATGCTCCACAACCTATTTGCCCTCTACCTGCTGGCTCATTAGCT
ACCACTGTCTACCCCCTTGCATCAGCCAGATTTTCGTCGCTATAGTGAAACACCTGGGGTAGGCATCTTT
ATAAAGAGAGGAGGGTTATGTGGGGCCGCTGTTCTTGAGGTGCAAGGTGTAGGGGCTGCATCTGGTGAGG
GCTGAAAGTAGCATGGGACATCACATGACAGGAGCCGGCCAGCATACTGGCAGCAGCATTTCCCACTTCC
CCTCTGCTCTTTGGTTCTGCTCTGAACTGGCTGTTGGATCTAGAACCGTGCTGTGGGGCTGGCATTGAGG
TGTAACGGGTGAAGCTGCCGTCTGCAATGCCAGTGTCCCATGTGGGTGCTGGTTCAAGTTCTGGCTGCTC
TACTTCCAGTCCAGCTTTGTGACATGGCCTGGGAAAGCAGTAGAAGATGGCGCATGTGCTTGGGCCCTTG
CACCTAGGTGGGAGGCCTGGAAGAAGCTCCTGGCATCCAGCTTTGGCCTGGCCCAGCTCTGGCCATTACA
GCCATCTGGGGAGTGAACCAGCAGGTGGAAGTACTGTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTC
CTAGTCTCCCTTTCAAATAAATAAATGTCTTTTAAAAATACACAAATAAATAAATAAATAAATAAAATTT
GTAACTTATCACCAACCTACAGCCAAGTTAAACAGGACTGTTAAAGACATCACAGGCTGGCGCCGCGGCT
CACTGGGCTAATCCTCCGCCTGCGGCACCGACACCCCAGGTTCTAGTCCCGGTTGGGGTGCTGGATTCTG
TCCTGGCTGCTCCTCTTCCAGTCCAGCTCTCTGCTGTGGCCCGGGAGTGCAGTGGAGGATGGCCCAGGTC
CTTGGGCCCTGCACCTGCATGGGAGACCGGGAGGGGGCACCTGGCTCCTGGCTTCGGATCTGCGCAGCAT
GCTGGCCGCAATGCACTGGCCGTGACGGCCACTTGGCGGGGTGAACCAACAGAAAATGGAGGACCTTTCT
CTCTGTCTTCTCTCTCACTGTCTGACTCTGCCTGTCCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
AAAAGAAAGAAAGAAAGAAAGAAAAGACATCACAGATGATCCCAAACTGTGGTCAGACTGGAACAGAACT
GCAGTGATCACAATCAGATAATCAGATTCACAATGGTCAAGTGTGTTCCCCAGGCCTCACTGGACCCAGG
GGCTTCTAGTGTGTGTGTGTGTGCGTGTGCGTGTGTGTTCACGTTATGCACATGTATGTGTGTGAGTACA
TGTTACACACATGTATATGTTGAGGGTCAGCCATGTCAGGCTGGGAGCAGGAAGTTTGACTGACGGCCTC
TCTCTCTGCAGCACGGAGCTGTCCAGAGCACAGCCACTATGAGTCCTGTGCCCGCGGCTGCCCGCTGAGC
TGCGGGGACCTCCCGGTGTCCGGGGGCTGTGGCTCAGAATGCTACGAGAGCTGCGTGTGCGATGAAGGCT
TTGTGCTCAGCGGCGAGTCCTGCGTGCCCCTGGCCTCCTGTGGCTGTGTATACCAGGGCTCCTACTACCA
GCCAGGCCAGACCTTCCACCCTGGCCCCGGGTGCGACTCCCTGTGCCACTGCCAGGAGGGTGGCCTGGTG
TCCTGTGAGGACTCCTCCTGCGGCCCGCAAGAGGAGTGCCAGCTGTCTGGTGGCGTCCTGGGCTGCGTGG
TTGTGGGCTCTGCCACGTGTCAGGCATCCGGAGACCCCCATTACGTCACATTCGATGGCAGCCGCTTTGA
CTTCATGGGCACCTGTGTGTACGTGCTGGCTCAGACCTGCGGGACCCCGCCCGGCCGGCCCCAGTTCACC
GTCCTGCAGGAGAACGAGGCCTGGGGCAACGGACAGGTCAGCGTGACCAAAGCAATCACGGTCCTCGTGG
GCAACCACACCATACGTCTGGAGCAGAGTCAGCAGAAGGTCACGGTGAGAACCACGGCTTGCATGGGAGG
GGCGGTGCTGGAGTGGGGGTGCGGCCTGATCTCCCCTGTCTGTGCCCCTCCACCTCTCCGCCTTCGGCCC
CTCGGCCTACGCTGCACTGTCCTGTCTGAAGGCCTCTTCCACCCCCTCCTCCCTCCTTCTCCATCCTCTC
GCCACCCCTCCCCTAACCCGTCCCAGCCCCCTCCTTCATCTCCCACCCTATCAGAGGTTGGGAAAGATCA
GAGAGTGGGCAGGGTCCAAGGACAGAAGATGGGGTGTGGGGGTGACCCTGTCCCCGACTCGACAACCACA
GTAGCTACAGACTTTGCAGGCTGAGCCCGGACCCTGCGTCGAGCCCACAGGTCACTCTGTCTTCCCTGTG
GCAGCTGGAGGCACCACGATTAACTCTCTTTTTGTAGATGAGGAGGTCCGGCTGTTGGCCTAAGAAACTG
GCATTTCGGCTGCACTGGAGGGGAGGGGACTTTCTCAGAGACCCCACTCCCACACAGCCACTGCTCTGGT
CCTGTCCTGGATCAGTAGATGACTTCATCACACATGACATAGCTTAATACAGCCCCCAACCCAGCCCAGC
TCTTAGAGCAGCGCCCCGACTGTGGTTTGCTCTAAGCTTCTTGTAATTGACATCTCACTACATCCCACCC
CACCCCCGCTTCCCTGAGAAGTAGGAAGTATCATCCCCACTATGCAGAGGAGCAAGGCGAGGCACAGGTG
AAGTCAGTTCTCTGCTTGGGAAGTCCAGCAGTTGGTTATGCTCTGTGGGGACAGACACAGCCAGGAACCC
GCCTTTCTCCTGCCTCCTCCCCTGATTCCTCACACTCCAAAAGACTCCAGCGTGCACCACATCTGCCCCC
ACGGAACGAGGAGCAAGTGGGTTAATCGGGGTGGGTTTGCACAGCTTTGCCAGTGGCCAGCAGCCATTGG
CTGAAACCACGGGTCCACAGCTCTCGGGGCTGAAGGTCATTTGCCCAGGAATGTAGCTTGGCCTTCCAGC
CTTCAGTGTTTAAAGCAAAAACCTTCAGTGGCCGCTCACCCGTTAAAATGAAGCCTTCCAGTCAGGGCTG
GTGATCACCTGGATGCACGGCAGCCCCCCCCCCCCCACGCCTCCCCGCTCCTTCACGTTCACACGGCCCT
CTGCTCTGTGTCTTCCGGCTTGTCACTGGATACAGGGCCCGCCCTAATCTCCTCTCCAGGTCATTCCCCT
AATTATGTCCATGAAGATTCTTATTCCAAAGTCACATTCTGCTGTTGAAAAAACAATATTTATTTATTTG
AAATAATTGCAGAGATAGAGAAAGAGAAAGAGAGAGAGAGAGAGAGAGAGAGAGATCTTCCATCCGCTGG
GTCATGCCCCCAAATACCCACAACAGCCAGAGCTGGGCCAGGCTGAAGCCAGGAGCCTGGAACTCCATCC
GGATCCTCCCACTTGGGTACAGGGGCCCAAGTACTTGGGCCATCCTCTGTTGCTTTCCCAGGCACTTAGC
AGGAAGCTGGATCAGAAGCAGAACAGCTGGGACTCAAACCAGTGCTCACAGGGGATGCCGGCGCATCACA
GGCAGTGGCTTAACATGCTGTGCCATGAGACGAGCCCCTAAAGTCACAGTCTGTGATTCTAAGTAGACAT
ATATTTTGGAGACCAGCATCCAACCCCCTATAAGACTCTATTATTATACCCATTTCACAAATACAGAAAC
CACAACTCAGGGAGGTGAGGCACCACAGACCCTCACTGTCGGGCAGGGGCTGTAGCGGGTGTGGCTGGGC
CGAACCCACCCTCTGAGACCCTGCTCCTCCCGCCTCCCAGGTGGACGGCGTGGACCTCAAGCCGCCGGTG
GAGCTGGACAAGGGCGAGATCCGCATCAACCAGCACGGCTCGGACGTGGTGATCGAGACCGACTTCGGCC
TGCGCGTGGTCTACGACCTCATGTACTACGTGCGGGTCACTGTCCCGGGCAACTACTACCAGCAGATGTG
TGGCCTCTGTGGTGATTACAACGGCAAGGCCGAGGACGACTTCCAGAAGCGCAACGGCTCACAGGCGGCC
AACCCCAACGACTTCGGCAACTCCTGGGAGGAGGCCGTGCCGGGCTCGCCCTGCCTGCCGCCCCCTGAGT
GCGACTCGGATGCAGATTGCAAGTGCACCCCCGAGCTGGAGGAGCAGTACAAGAAGGAGGAGTTCTGTGG
GCTGCTCACCAGCTCCACGGGCCCGCTGTCCTCCTGCCACAAGCTGCTGGATCCTCAGAACTCCTTGCAA
GACTGTGTCTTCGACCTCTGCCTGGGCGGCGGGAACGAGAGCATCTTGTGCGGCCACATCCACGCCTACG
TGAGCGCCTGCCAGGCAGCTGGAGGCCAGGTTCAGCCCTGGAGGAATGAAAAGTTCTGTCGTGAGTGAGG
GGGACCCCGCAGGGCAGGGGCAGGGACATTGAGACCCAGACGGTGGGGCTCGTGCCGTGCTGCCCTTGCA
GGGCTCTGACGGGTGCCCCCTCCTTGCAGCGATGGAGTGCCCCCCTCACAGCCACTACGAGCTGTGCGCA
GACACCTGCTCCCTGGGCTGCTCGGCTCTCAGCGGCTCCGTGCAGTGCCCGGATGGCTGTGCTGAGGGCT
GCCAGTGTGACCCTGACTTCTACTACAACGGTGAAGCCTGCGTGCCCATCCAGCAGTGTGGCTGCTACCA
CAACGGTGTCTACTACGAGGTAGGATCGCGGACACCCCCCCTCCCTGGAGGGCCGCTCCTGGGACCTCGT
CCTGAGCCCCCTCCCACCACCGCTCTTCCCCCAGTCTGTCCACTCCTGCCCACGCCTCTCGCTTGCCCAG
CTCCTTTTCCCCACCTGGATCTCTCCTACCCCCATTTTCCGGCCTCTGGATGACCATCCATTTGCCAGTG
GGGGAGCTCTTTCTAAATATGACTGTGGGCCCTGGGATACCCTTGCTGCCCACATCAAGTTGGGGCTTTG
GGGCTCAGGCGTGGACAAAGGTGGCTCTTAGGACCACAAGGATGAGATTTGGTCTTTATCCCACCTGGCC
AACTCCGCGTCCCTCGTCCACTCTCTCACTGTCGGTCCTTTTTAAAAAATGTTTTTATATATTTATTTTC
ATTTTATATGAAAGGCAGAAAGAGAAACAGAGAGAAATCATCCATCCCCCTCCCCCTCCCCCTCTCCCTC
TCTGTCTCCCTCTCCCTCTTCCTCTCTCTCTCCCTCTTCCTCTCTCTCTCCGTCTCTCTCCCTCTCTCTC
TCTCTCTTTCTCTCTTTCTCATCCATCCACTGGTTCACTTCCCAAATGTCCACAACAGCCGCCAGAGCCA
GGAGCCCAGAACTCCATCCAGGTCTCCTGAGTACTGGAGCCGGCACCTGTGGCCTCCCAGGGTGCACATT
AGCAGGGAGCCAGACTGGAAGCGGAGGAGCCGGGACTTGAAGCAGGCAACTCTGATACGGGATGCAGGCG
CCCCCGGCAGCCGCATAGACCCCGCTGGGCAAACACCCAGCCCTGTGTTAGTCCTCATTTCAGCTGAGTA
GAGTTTGTCCGACGTGCATCTCCCTCACAGGGCTGTGAGCTCTACGAGGGCAGGGGCCTCTCTGTTCTCC
TCCCATGCATAAGAGCAGTAACTCGTCCACAGTAAGCAGGAAACACGCGGATGGTGCTTCCTGGGTGCCC
TTGCTGTGTGTATGTTTTACACGCACTGCCTATGGACGTAAGCTTTTTATTATGACGATGTTCAAACTTG
CGAAAGTTCAGAGAACAATAGAACGAACCTCCACGTGTCTGAGCATCCCTGGGGCTGCCAAAGTTGACGT
CTTCCCCAGTCGTGTTCTCTAGCCTCCTTCCATCTCCTACTGGGATGATTGGCCGGAAGCCTCTTTTCAT
CCGACGTATACTTTCTATGTGCTGCTAACTCAGGCTTGCACGGGTAGGTGCTGTTGTTATCTGCAGATTG
CAGGTGAGGAGACTGAGGCAGAACAAGGCTAAGTAATTTGCCCAGAGTATCATAGTCTGGAGACAGCAGA
GACGGACTTCAGATGGCCCGCTCCGAATTCCCGCCACACTCTTTCTCCCACACTGCAGAGCTGAATGAAT
GAGTGGAAGTGAATGTATGCATTGGGTCCCAAGGCCATGCTGTCCACAGGGTCCCTGATGGCCATGCCTT
ATGTCCCTGCAGCCGGAGCAGACGGTACTGACTGACAACTGCCAGCAGCAGTGCGTGTGCCACTCAGGCA
CAGGCGTGACGTGTCAGGAACACAGCTGCAAGCCGGGGCAGGTGTGCGAGCCCTCGGGAGGCGTCCTGAG
CTGCGTCTCCAAAGGTACTGGGCTGGGGCTGGGGCTGGGACTGGCGAGGCACAGGGGACACGGGGCCAGG
AGCCAAGAGGGGCGTGGCAGCGGTGTACATGGTGGGAGGTGCAGAAGACTGAGGATGAGGTCTCAGGCCC
TGCATTGATCTTTCAGGGCCAGCAGCAGAGACTCCCTTGTCTTGGGGACCTTCCAGGCCAGGTTTCCCAG
AGCACATGTCAGAGACCCTCAGTGAAAGGCAAATCTGGAGGCGCCTCCAAGGCCATGAGGCTGAAACACA
GGGGCCCCAGGAAGCATGTTGGGACCCCTGCCACTGGTTCTTCCCAGGAAGTCCTCTAGGACCTTGGTGT
TGGGCACAGAGAGCCCTAAGACGTGGGTCTCAGATGCAGAGAAGCCCACAGGGCTAGGACCTCAGCTATG
GAGGAAGCTAGGGCCCAGGTCCCATATGGACACCCTGAGGCCGGCATCGCAGACACAGGCGCCCTCAGGG
CTGGCATCGCAAACCCAAGGACTCCTGAGGTTCAGGTCTCAACCTAGGTCCCCTGCAAAGAAAACCCTGG
GGACCCCGGGAGCTGGGTCTCAGATGTGGCAACTCTGGATGCTCATGTCTTAGACACAGCCTGGCCCAGG
GCCTGAGTGCCAGACCTGGAGGACTCCCTGGGCCTGGTCTTAGAAACAGCATCTGAGCCAGCGCCATGGC
TCACTAGGCTAATCCTCCGCCTAGCGGCGCCGGCACACCGGGTTCTAGTCCTGGTTGGGGTGCCGGATTC
TGTCCCGGTTGCCCCTCTTCCAGCCCAGCTCTCTGCTGTGGCCAGGGAGTGCAGTGGAGGATGGCGCAGG
TGCTTGGGCTCTGCACCCCATGGGAGACCAGGAAAAGCACCTGGCTCCTGGCTTCTGCCATCGGATCAGC
GCGGTGCGCCGGCCGCAGCGCGCTGGCTGCGGCGGCCATTGGAGGGTGAACCAACGGCAAAGGAAGACCT
TTCTCTCTGTCTCTCTCTCACTGTCCACTCTGCCTGTCAAAAAAAAAAAAAAAAAAAATATAAAGAAAGA
AAGAACAGCATCTGTGGGATCTTAGAAGTAAACCTTGGATCTGCTCTCTCCTGTGGTAGTCCTCGGGGCC
GGTGTCCCTGGCACAGCCTGAGGTCTTCTCTCCTCTGCCCCCAGACCCATGCCAGGGTGTCACGTGCCGG
CCCCAGGAGACCTGCAAGGAGCAGGACGGCCAGGGTGTGTGTGTGCCCAACTACAACGCCACGTGCTGGC
TGTGGGGCGACCCACACTACAACACCTTCGACGGCCGGGCCTTTGACTTCCAAGGCACCTGCAACTACAT
TCTGGTGCAAGTTGGCTGCCCGGGGGTGGACGCCGAGGGCCTGACACCCTTTACCATCACCACCAAGAAT
GAGAACCGGGGCAACCCCACCGTGTCCTATGTGAAGGAGGTCACCGTGTCTGTCCTGGGCACCAACATCT
CCATCCACAAGGGCGAGACGGGCAAAGTCAGGGTATGTGTGGCAAGGTGGTCCCCTGAACCCCCAGGAGG
GCTGGGGGAGCCCTGACAGCCCCAGTGAGGAGCCGAGGTTCTCTGTATTCCCCTAGGCCTGAACCAAATC
AGCCACCGAACAAACAGGCTTAGCTCTGTGCTGAGCACTTCGGCATCTGCCTCTCAGTAGACTTGGGTTT
GAATCCAAGCTCTGCTTCCCAGGTGCTGGTGGGTGGGCCATGTGCCCAGCTCAGAGTCTGGCCCAGATGA
GCTCCAGAGAGCAGATTAAGAGGCCTGGAGACTCGGACCCAGAGGGAAATCCAGGGAGGTGAGGGCAGAG
AGGTGGGCCTCTGCTTCTGCCCAGGGAAGAGCCATGTGGGTCCACTCAGCAGACACTAAGATGGGCATGC
GCACAGGAGTGCACAGAGAGTGTGTACGGCCAGTGGGGAGAGCCAGAGACAGAGAGGACAGTGACGGGAG
GGAGTGCAGAGACCCAAAGAAAGAGGGAGAAGCGGAGGTCCAGCAAGAGAGCTAGACCCAGGTGAAGCAA
CAGAGAGGGAGGAGAGAGCAATCCAGAGGAAGAAGCAGACACAAAGACGGGAACAGATGTTCATGACATT
GTCAAACGGCTATACAACTTTCAGGCGGGGGGAAAGCCATGGCAGCTCCCAACCCCCGTCAGGACAAGGT
GGGGAGTCGAGAGCTGGGGCCTCCCACCCTGAGTGACCCAGGCACCTGCTTCCCCCTCCGCCAGGTGAAC
GGCGTGCTGACGACGCTGCCTGTCTCCGCGGCCGGCGGGCGGATTTACGTGTCCCATGGAGTGTGGAAGG
CTGTGCTGGTGACAGACTTTGGGCTGCAGGTCACCTATGACTGGGACAATCGCGTGGAGGTGATCCTGCC
CAGCAGCTATCACGGCGCCGTGTGCGGGCTCTGTGGAAACCTGGATGGCGACCCCAACAATGACCAAGTC
TTTCCCAACGGCACAGCCGCTCCCTCCATACCCGCCTGGGGTGGCAGCTGGCGGGCCCCAGGCTGGGACC
CACTGTGCTGGGACGAGTGCCAGGGCTCCTGCCCGACGTGTTCTGAGGACCTGCTGGAGAAGTACCAGGG
CCCTGGCTTCTGCGGCCCCCTGGTCCCCGGGACCGAGAGCCCCTTCGCCGACTGCCTCGCCCACGTGTCG
CCCGACAGCTACTTCCAGGGCTGTGTCCTGGACGTGTGCCTGAGTGGTGGGGCTCAGGACATACTTTGCC
AGGCGCTGGCTAGCTATGCGGCTGCCTGCCAGGCCGCTGGCGTCGCCATCCAGGAATGGAGGGAGCAGGC
CGGCTGTGGTGAGTGCCGGGGAGGGGGCAGAGGCTGGGGGTGGGGTCCTGTCTGCCAGGCGGGCTTGACT
GCTCTCCAGCCCTTCTGGGTTCTTCTTGTATTTCTGTTCCCGAGCGCCTGTCTCTCGGCGCTTCTCTGTG
TGTCTCCCTTTGGGTCTCTCTGTATCCCTCTCTGTGCCTCTTGCTCTCTCTCCTTCGGTCTCTGTCTCTC
TTCCTCTCTCTGTCAGCCCCTTGGTTTCTGTATTTTGCCACCGTGTCCTTGGACCCCACCCCTAACATCC
TTCCCTCTTTCTCCCTCTCCCACCCGCCCGCAGGGGTCTCCTGCCCTGAAAACAGCCACTACGAGCTCTG
TGGCTCACCCTGCCCAGCCAGCTGCCCGGCTCCTGAGTCCACCAAGACCTCAGCCCTCTGTGAAGGTTCC
TGCGTGGAGGGCTGCCAGTGCGACACGGGCTTCGTGTTGAGCGCTGGGCGCTGCGTGCCCCTGGAGGGCG
GCTGCGGCTGCTGGGCCAACAGCACCTACTATGAGACCGGCAGCGAGTTCTGGACGGACGCCACATGCTC
CCAGCGGTGCCGCTGTGGGGCTGAGGGCGGCTCGCTGGTCTGCACACCTGCCAGCTGCGCCAAGGGCGAG
GCATGTGCCCTGCTGTCCTCGGGCCAGTACGGCTGCCAACCAGTGAGCACTGCGGAGTGCCAGGCCTGGG
GTGACCCCCATTACGTCACCCTGGACGGGCACCGATTCGACTTCCAAGGCACCTGCGAGTACCTGCTGAG
TGCACCCTGCTCTGAGCCGCCGGCCGGGACTGAGAATTTCACCGTCACCGTTGTCAACGAGCACCGGGGC
AGCCAGGCTGTCAGCTACACCCGCAGTGTCACCCTGTATATCTACGACCACCGCCTGACCCTCAGCGCCC
GCTGGCCTCGGCAGCTACAGGTGAGGAGGGCATGGGCCGCACAGGAGCAGGTGCCGGCCCTGGGGTTGCC
TGGGACGGTAAGAGGTCCGCCAGGTCCCTGAACACCTTACAGCTTAGCTGGGACTTGATGGACGGGGTCT
GGAGCATAGGCCTGGGAGCTGGATGGTGTGAGGCAGTGGCTCCCCCTCCCCCTGCGCTGAGCCTTAGTTT
ATTCCTACGAGAAATGGGCCCAAGAAGTCATGTGCTGCAGGGTGGCGTCAAGAGCCTGTTGTTTTTACTA
CAAGAAGCCCTGGACTCAGAGTCCCACAGCTCCTAGGTGGTGGAGACGGGCTTCGAACCTGACTGGCCCG
CCCCGCCCTGCCCCACGCCCGCAGGTGGACGGCGAGTTCGTGACGCTGCCCTTCCAGCTGGACTCGCGCC
TGAACGCGTATGTGAGCAGCGCCGACGTGGTGGTGACCACGGCCGCCGGGATCTCGCTCAGTTTCGACGG
GGACAGCTTCGTGCGCCTCCGCGTGCCGGCGGCCTACGCGGGCGCCCTCTGCGGCCTGTGCGGGGACTAC
AACGAGAACTCGGCCGACGACCTGGCAGCGGTGAACGGGAAGCCCGAGGGCTGGCAGGTGGGCGGTGAGC
CGGGCTGCGAAAAGTGCGAGCCCGAGCCGTGCCCGGAGCCGTGCACGCCCGAGGAGCAGGAGCCCTTCGG
CGGCCCCGACGCCTGCGGGGTGATCATGTCCCGCAAAGGCCCGCTGGGGCCCTGCCACTGCATCGTGCCG
CCGGAGCAGTACTTCCAGGCCTGCCTGCTGGACGCCTGCCAGGCCCAGGGCCATCCGGGAGGCTTCTGCC
CTGCGGTGGCCGCCTATGTGGCTGCCTGTCAGGCCGCCGGGGCCGAGATCGGCGAATGGAGGCTGCCGGA
CTTCTGTTGTGAGTACTGGCCCTTGCACGTTCACAAAGCAGGCGCCTCCTCCTCTCCCCAGCTTCTTCCC
TCTTCCTGGGCTCAGGGACCAGCGAGGTGTCTGTTAGACTATGCGCGGCGACAGACTGGTCAGGTACACC
TACGGTCAGGGCTCTGAGGACCCCAAACAGCTGCCTAAGACAGGGTCCGGGAGTCCCTGCAGTTCCTTTT
GGACAGATAGATCTCCTCGGTCAGGTGCATCCTGGCAGCTTCAACAACCAGACCTAGTTGTAGTCCCCGC
ACAGGCCGCAGAGGGCGCCCGCGTAGGCCGCCGGCACGCGGAGGCGCACGAAGCTGTCCCCGTCGAAACT
CTGTGGGTCAGAGTCACACCCTAAAACAAACGGAGTCGGGGAGACTCCAGCAGGTGCTCGGACACCCACG
CCAGGCACAGCCCCACAGGCGCACGCCCCGGGATACCTGTGGCTGATCCTAACCCTTGTATCCTGGTCAC
ACATCCATGACCAGACACTCACTGGACACTTAGAAGTTTTAGGGTGTGACACACACCTGGTCAGAGGGTG
TTGCCCCCACCCCGTGGGAGGTGCTGGGTCCGATCCTGGTACCTGGGTGTGTTGTCCCAGGGGAAGCCTC
ACCCTCGTCCCGTCTCACCCCCGCAGCCATCCAGTGTCCAGCCAACAGCCACTACGAGCTCTGCGGCAAC
TCCTGCCCCGTGAGCTGCCCCAGCCTCTCGGCGCCCGAAGGCTGCGAGCCAACCTGCCGCGAAGGTTGCT
ACTGCGACGCCGGCTACGTGCTCAGCGGTGACACATGCGTCCCCCTGGGCCAGTGCGGCTGCCTCCACGA
GGGCCGCTACTACCCACTGGGCGCAGCCTTCTACCCAGGGCCCGAGTGCGAGAGACGCTGTGAGTGCGGC
CCAGGGGGCCAAGTCAGCTGCAAGGAGGGTGCGGCCTGCGGGCCTTACGAGGAGTGCCGGCTGGAGGACG
GCGTCCAGGCCTGTCACCCCAAAGGCTGCGGCCGCTGCCTGGCCGACGGAGGCATCCACTACCTCACCCT
GGACGGACGCGTCTACGACCTGCACGGCTCCTGCTCCTACATCTTGGCCCAGGTCTGTGACCCGCAGCCC
GGGGATGAGGACTTCACCATCGTCCTGGAGAAGAACGCGGCTGGAGAACCGCAGCGCGTGCTGGTTACCG
TGGCTGGCCAGGTCGTGGGCCTGGCCCGGGGACCGGAGGTGAGTGGCCCGCCCAGGGACCCAGGGAGACA
CAGCCAAGTCCTGGGCTTCTGGAGGTTCTGTGAGCCACTCACCCGCCCCTTCCTCCCGTGCAGGTCACCA
TCGACGGCGAGGTGGTGGACCTGCCCGTGGCCGTGGGCCATGTGCGGGTGACCTCCGAGGGCCGAAACCT
GATCCTGCAGACAACCGAGGGGCTGCGGCTTCTCTTTGACGGCGACGCCCACGTCCTCATCTCCATCCCC
AGCCCCTTCCACGGTCGGCTCTGCGGCCTGTGCGGCAACTTCAACGGCAACTGGAGCGACGACTTCGAGC
TACCCGACGGCTCGGTGGCCCCCAGCGTGGAGGACTTCGGGGCGGCGTGGCGGGCACCCGGCTCCCCCCA
GGACTGCACCGAGGGCTGTGGGGCCCAAGGCTGCCCCGTGTGCCGGGCGGAGGAGACGGCACCCTACGAG
AGCAGCGAGCTCTGTGGGCGGCTGCTCGACCCCAGCGGCCCCTGGGCCGCCTGCCACGAGGTGCTGAGTC
CTTCCGAGTACTTCCGCCAGTGCGTGTACGACCTGTGCGCCCAGAAGGGCGACCAGGCCTTCCTCTGCCG
CAGCCTGGCCGCCTACACAGCCGACTGTCAGGCGGCCGGCGTGGCCGTGAAGCCCTGGAGGACAGACAGC
TTCTGCCGTGAGTGTCGGGGGCGGGGGTGGGGTGGGGTGGTGCCTGGGAGGTCTCCCGCCCGCCACGCCC
CGCCCTCCTCCGTCTGCATCTCAGGAGCAGCCCAGCCCCCTCCTGCGAAAGCTCCTCCCACGCCCGCTGC
TCGCCTGGCGGCTCCCCTCTGCCCCCTCTGACCGCCCCCTTTCTCCTCAAAGCGCTTCACATCTTCCCTG
GGCCTCCATCCTGGGCACTTTCTCACCTTGCAGCCTGAGTCTGTGGCTTGCCAGCCCCCTGGGGGCAACA
CCGGCTCCCCTCAGAGCACCCAGGACAAAGCTGCAGGCCGTTGGGCCCCCAAACCAGGGCTGTCCCTCTC
CCCCAAATCTCCTGCTGCAAGGACATGCCCCTCCCCAACCTAGGCTCTGCAGGAAAAGGCCATGGCCGAG
TTTGCTTCGGTCTCCTCAGAGCAGGACCCCTACCCAGGGCCTTATAGGCTTCACAGCCTTGGCCTTGTCC
TCAGCGGGGACAGTGGAGCCTGTCTCCTGCTCCGAGCTCTCTCCTGACTGCCAGCCCCAGGGGTCCAGTG
TGAGCGCGTTCCCTCCAGCATCCTCACAATGTCTTGCACCTGCTGTGTTCCCAGACGACATCAGCGCCTC
ACCCCAAGCCAGTCCTCCTTCCCCTGGCCCCCCCGTGTCCCGGCACTGGGCAGCCACAGCAGCCACCAGT
TCTCAGCCAGAGCCCAGGACCTCACCCCGAATGCTTCCCCTGGGTCCTCCCAGCCCCCTGTGTTTCACTT
GCCCCGGCCCTGAGCCCCAAGCCTGACCTTGCCCAATGCAGCACAGCCACATGGGCCCCTCATGGCCTGT
CCCTCCGGTTTGAGCTCTGCAGGGCAGAGTTTGGGGCTGTCTGCGTCATTTGGGTCATCAGCGCAGCCCA
CGCCCGGGCTGGCTGTATAGAAGCTCTTGTGGCCGTCCATGGAGGCGTGACCCCAGCCCGCCATGTCTCT
CTCCACAGCGCTGCAGTGCCCAGCTAACAGCCACTACTCCGTCTGCACCTACTCCTGCCAGGGCTCCTGC
ACAGCGCTCTCTGGCCTCACGGGCTGCACCAGCCACTGCTTCGAGGGCTGTGAGTGTGACGACCGCTTCC
TGCTCGCCCAGGGCGTCTGCGTCCCCTCCCAGGAGTGTGGCTGCACCTACAATGGCCGGTACTTGCCTGT
GAGTAGGGGCTCGAGGTGGGCAGGGAGGGCGGGAGGGAGTCTAACCTCCCCGAGGACTGGGACAGGGGTG
CCGCTGCCATCTCCCAGCTGGGACACTTGCTGTGTGTGATGCTGCTGACCTCCAACCTGGTAGTCTAACA
CGGAAGCAATGTGTTCTCCCACCATTCCGGAAATCAGTACCCCCACGCTGGAGGCAGGCTGCCCCCAGCC
CCCCCCCCCCCCGCAGTCTAACTGCTCCCTGCAGCTTCTGGGGGCTGCCAGCATCCCGTGCCTTGGGGCT
GCATCCCTGCAGTATCGGCCTGTGTGTGTCAGGAACGCTCCCTCCAGCTTCCTCTTGGGAGGACACCCGG
ATAATCCAGCATGAATTCTCCACTTCCAAGTCCTCGGTTTAACACCGCTGCAAACACCTTTTTCCCACAG
AAGGGGATGTTTACTTGGTCCCAGGGATTGGGATCTGATGTCTGTGAGGGACCACACTTGGCCTGCCATG
GTCCCCACCCTGAACCTCATTTTCTCCACCTGTAAGATGGGGGTAGTCATTGTCCTTGCACCACAGGTGT
GTAAGGGAAGGTGTGATTGTGTGGAAGACGCCAGCTGTGCCCTGGGCCTGGCACTCCCTGCGCATCCCAC
TCAGGGAAAGGAATAGGAACTTAGACGGTGCTCTAGGCAGCCCCCAACCATCACTTACTCTGCAAACTGA
AATGCAATCCTTGATTTATTCGTGTTAACAGTATTTTTTTGGGCTCTTGCTCTGCACTGGTTGTTATGGG
TGTTTAAAGATTTAGTTATTTATTCGAAGAGGAGAGAGAGAGAGAGATTTTTCATCTGCTGGTTCACTCC
CCAAAGGACCACAAGACTGGGACTGGTCCAGCTCAAAACCAAGAGCCTGGAGCTCCATCCTGGTCTCCCA
CATGGGAGGCAGGGCCCCAACTACCTGGGCCGTCCTCTGCTGCCTTCCCAGGTGCATTTAGGGAGCTGGA
TCAGAAGTGGAGCTGCTGGGACTTATTGGCGCTATTGCTGTTTCCCCCTGTTACAGAGCCATCCCTGTAA
CCCCCCAACCTCTTACCTGCCCCCTTCTGCACACAGGTGAACTCCTCCCTCCTGTCCTCAGACTGCAACG
AGCTCTGCTACTGCTCCCCAGACGCAGACCTGCAGTGCTATGAGGCCAGCTGCAAGGCGGGCAGCGTCTG
CGAGGTCCAGAACGGGGTTCGGGTCTGCCAGGCCTCCGGCGCCCAGTGTACCCTCTCCGCAGGCTCCAAC
CTCACCACCTTTGATGGGGCCCTCAGTGCCTTCAACTCCTCTGGTGTCTATGAGCTCTCCTTCCGCTGCC
CAGGACTACAGACCCCCTGGTATCGTGTGGTGGCAGCCATCCAGCCCTGCGCTGGCACAGCTGAGACGGT
GGGCCAGCTCCACGTCTTCTTCGAGGACGGGATAGTCACCGTGACCCGAGACGGGGGCGTGTGGGTAAGC
CTGGGCGTGGCGGCGGGCGGGCGGGGGTCGGGGTGATGTGCAGTGTCCTTGCATTGTTGCCTGCCTGTCT
TGCTGTCTCTTTCCTCTGCAATTTGATCTCTGGATTGGTATTTGCCTCAGTATTTCAGCTGAGTTCGTGG
GAGGCAGAACGCATTCAAGGAACGTCATAGTTTTGATATTAGCGTCAATCAGGACTCCATTGGTTGCAGG
TGACAGAAAAACCGAGCTCAAACTTCCTGAGAAGCAGAAGTGAATTGATTGGCCCACGGAACTAGAAAAC
TCAGGGTCAGGACTATAGGCCTCTATGTTTTAGATGTGCTGGTCACCAAGTCGGCTTGATTCTCCGGCTT
TCCTCTCGCAGACCCACACCGCACTGGTTTAGCTACGCTGGACCAGTTTGGGCCCTTGCCTATCCCTGGA
ACCAGTCACCATGGCCTTGGGGATGGAATGCTGTGATTGGTCAGGTGGTTTTTATTGGAGGGGGTAGGGC
TGGTCACCCAGGCAGCTCTGCCTAGCACATGCCAAACGTCAGGCTCCCGGAAGGAAGCTGCGGAAGCTGC
ACCTTGCATCAGCTCCGGAACCTGAGCCCTGGGTGCGCAGGGATCCTGTCTGTCCCCGTCACCCCTTTAT
TCCCAGTCCCCAGATCCAGGCCTGGCACCTGGACCCCGGAAAAGTCGGGTTTGGTGACTCAGCTCAGCCC
CGCCCCTCTGAGTCAGGCTGGGAGCCGACTCCCACTCCAGGGCCGACTCGAACCCAGTTCTCATACTCAG
GAAGCAGCCGGGAGAGGCTTCCCCTGCCCCGCCCCGCCCCTCCTGGCTGTCTCCATCCTCCCTCCCTTTC
TCCCTCCTCCCTTCTTTCCTCCCTCCCCCCCATCTCAGCTCCACACCTACTTTTCCCATCAGTCTGCCTG
TCTCTCCAGTCACCCGACAGAGGCAGACTCAGATACAAAAGCATTTCGGGAAAAAAAGAGAACTTCTGTT
TAAAAGCAATTTTTGCTAGAGAGACTAACTTTAGAAGGATGAACTTACCAACAAAAACAAACAACACTTG
TTTCAAGGGAAAAAAAACTCCCGACGTCGCTGAGTCTGCAACCCAGGACGAGGAAGCAGGCATCTCCTCA
CAGGCCTATTATATTTGCTATGCTCACTACTTATCTTTAGACACCAGCATTACAATGATAAGGGATAAAC
TCTCCTCTTGGGGAAGCAGTAACAGAGAACATGCTCCTCAGAGGCCTTTTGTTTTGAAAAGTAGACCCCA
ACAGGGTGGCCCTGAGGTTGGTTGGTTGGGATGGGCCTCTGGTACAAAGTCCCTCCCACCCCCGACCCTG
GGACACTGCGTGGGTCCCCCAATCCCTTTCTCCATCCGCCCAGTCTCCAGCTATCTCCCCGAGTCTGTCT
GGCCACACGCAGCCTCCATCCGGGTCTTCATGGCTTTCAGAACCTGCTCCTTGCTTTAGTTCCTGAACTC
ATGACCTCGCTCGGGGTCTCCGTCCTGGTAACTCGCCGGCTGCGCTGCGGCTCCGGCCTCACCTGTGTCT
GCCCGCCTCCATCGCCAGGTGAATGGTCGCCAGGTGGACCTCCCAGCTGAAGTGTTGACATCGGTGTCCG
TGACGCGCAACAGCGACAACTCCATACTTGTCCACCAAAAGGATGGGCTTTGGGTGTGGCTGGGCGCCGA
TGGGCAGCTGACCGTGACGGTGGACAGTGGCCGCGCTGGCCAGCTGTGTGGGGCCTGCGGAAACTTTGAC
GGAGACCCGACCAACGACAAGTCGGTCACGGAGAGCTGGACAGCACCGGACCTTTCCCCATGGTGAGGAT
GTGGCGGGGAAGCCGGGCTTCTCGGGCGCGAGGGCGTTGGATGCTCACGGGAACGGGCAGTGGACGCGTG
GGCAAGAGGAGGTGCTGAGTGTCTGTCTCTTGCAGTGAGGACTG

Molecule Role : Vaximmutor
Related Vaccine(s): Vibrio cholerae hemA mutant vaccine

IV. Vaccine Information

1. Dukoral

a. Product Name:

Oral Inactivated Cholera vaccine

b. Tradename:

Dukoral

c. Manufacturer:

SBL Vaccine, Sanofi Pasteur Ltd

d. Vaccine Ontology ID:

VO_0010710

e. Type:

Subunit vaccine + Inactivated or "killed" vaccine

f. Status:

Licensed

g. Location Licensed:

Canada

h. Host Species for Licensed Use:

Human

i. Immunization Route

Oral

j. Storage

Store in a refrigerator (2°C – 8°C)

k . Approved Age for Licensed Use

2 months and older.

l. Description

Products: Proteins + killed bacteria. Other components: Saccharin.

2. Mutacol

a. Tradename:

Mutacol

b. Manufacturer:

Berna Biotech

c. Vaccine Ontology ID:

VO_0010728

d. Type:

Live, attenuated vaccine

e. Status:

Licensed

f. Location Licensed:

Canada

g. Host Species for Licensed Use:

Human

h. Immunization Route

oral

i . Approved Age for Licensed Use

Ages 2 and older

j. Description

Products: Live bacteria. It uses the V. Cholera live attenuated strain CVD 103-HGR. Other components: Yeast extract, Lactose, Aspartame.

3. V. cholerae CTB DNA Vaccine

a. Vaccine Ontology ID:

VO_0011360

b. Type:

DNA vaccine

c. Status:

Research

d. Antigen

cholera toxin B subunit (CTB) (Xu et al., 2009).

e. Gene Engineering of ctxB

Type: DNA vaccine construction
Description:
Detailed Gene Information: Click here.

f. Vector:

pJW4303 (Xu et al., 2009)

g. Immunization Route

Intramuscular injection (i.m.)

h. Mouse Response

Host Strain: BALB/c mice
Vaccination Protocol: Mice were immunized intramuscularly with 100 μg of the CTB DNA vaccine or vector control. The DNA immunizations for mice were given at Weeks 0 and 2 in DNA alone prime boost groups, or Week 0 only for those receiving DNA prime followed by KWC-B vaccine boost at Week 2 (Xu et al., 2009).
Challenge Protocol: Naïve suckling BALB/C mice at 5- to 6-day-old were separated from their mothers for 4 h, then inoculated by oral gavage with 50 μl of the appropriate serial dilution of the inoculum. To prepare the inoculum, V. cholerae O1 El Tor biotype Ogawa strain V48-65 growing overnight on an LB plate, were re-suspended in PBS and adjusted to 5 LD50 in 25 μl, then incubated with 25 of properly diluted immune or normal rabbit serum samples for 1 h at 37 °C. The suckling mice were kept in a 25 °C environment and observed every 4 h for 48 h post-inoculation. The survival rates were documented at each time point (Xu et al., 2009).
Efficacy: The immune sera elicited by CTB DNA vaccine were functional and protective against V. cholerae O1 infection in mice (Xu et al., 2009).

4. Vaxchora

a. Tradename:

Vaxchora

b. Manufacturer:

PaxVax, Inc.

c. CDC CVX code:

174

d. Type:

Live, attenuated vaccine

e. Status:

Licensed

f. Location Licensed:

USA

g. Host Species for Licensed Use:

Human

h. Immunization Route

Oral

i. Storage

Store at −25°C to −15°C (−13°F to 5°F), and protect from light and moisture.

j . Approved Age for Licensed Use

18 to 64 years of age.

k. Description

VAXCHORA (Cholera Vaccine, Live, Oral) is a live, attenuated bacterial vaccine suspension for oral administration containing the V. cholerae strain CVD 103-HgR. CVD 103-HgR was constructed from the serogroup O1 classical Inaba strain 569B by deleting the catalytic domain sequence of both copies of the ctxA gene, which prevents the synthesis of active cholera toxin (CT). (FDA: Vaxchora)

5. Vibrio cholerae hemA mutant vaccine

a. Vaccine Ontology ID:

VO_0002934

b. Type:

Live, attenuated vaccine

c. Status:

Research

d. Host Species as Laboratory Animal Model:

Rabbit

e. Gene Engineering of hemA

Type: Gene mutation
Description: This hemA mutant is from Vibrio cholerae (Ravichandran et al., 2006).
Detailed Gene Information: Click here.

f. Immunization Route

Oral immunization

g. Rabbit Response

Persistence: A hemA mutant is highly attenuated in rabbits (Ravichandran et al., 2006).
Efficacy: A hemA mutant induces protection in rabbits from challenge with wild type V. cholerae (Ravichandran et al., 2006).
Host Gene Response of IgA
- Gene Response: VCUSM2 elicited high titer of anti-LPS IgG/IgA antibodies on the fourth week post-vaccination. These titers were comparable to those elicited by the wild type strain and were significantly higher than titers pre-vaccination (Ravichandran et al., 2006).
- Detailed Gene Information: Click here.
Host Gene Response of IgG Fc fragment
- Gene Response: VCUSM2 elicited high titer of anti-LPS IgG/IgA antibodies on the fourth week post-vaccination. These titers were comparable to those elicited by the wild type strain and were significantly higher than titers pre-vaccination (Ravichandran et al., 2006).
- Detailed Gene Information: Click here.

V. References

1. Aktar et al., 2016: Aktar A, Rahman MA, Afrin S, Faruk MO, Uddin T, Akter A, Sami MIN, Yasmin T, Chowdhury F, Khan AI, Leung DT, LaRocque RC, Charles RC, Bhuiyan TR, Mandlik A, Kelly M, Kov�? P, Xu P, Calderwood SB, Harris JB, Qadri F, Ryan ET. O-Specific Polysaccharide-Specific Memory B Cell Responses in Young Children, Older Children, and Adults Infected with Vibrio cholerae O1 Ogawa in Bangladesh. Clinical and vaccine immunology : CVI. 2016; 23(5); 427-435. [PubMed: 27009211].

2. Eko et al., 2000: Eko FO, Mayr UB, Attridge SR, Lubitz W. Characterization and immunogenicity of Vibrio cholerae ghosts expressing toxin-coregulated pili. Journal of biotechnology. 2000; 83(1-2); 115-123. [PubMed: 11000467].

3. Ekong et al., 2009: Ekong EE, Okenu DN, Mania-Pramanik J, He Q, Igietseme JU, Ananaba GA, Lyn D, Black C, Eko FO. A Vibrio cholerae ghost-based subunit vaccine induces cross-protective chlamydial immunity that is enhanced by CTA2B, the nontoxic derivative of cholera toxin. FEMS immunology and medical microbiology. 2009; 55(2); 280-291. [PubMed: 19040663].

4. FDA: Vaxchora: FDA: Vaxchora vaccine information [https://www.fda.gov/downloads/BiologicsBloodVaccines/Vaccines/ApprovedProducts/UCM506235.pdf]

5. Kirn and Taylor, 2005: Kirn TJ, Taylor RK. TcpF is a soluble colonization factor and protective antigen secreted by El Tor and classical O1 and O139 Vibrio cholerae serogroups. Infection and immunity. 2005; 73(8); 4461-4470. [PubMed: 16040956].

6. Osek et al., 1994: Osek J, Jonson G, Svennerholm AM, Holmgren J. Role of antibodies against biotype-specific Vibrio cholerae pili in protection against experimental classical and El Tor cholera. Infection and immunity. 1994; 62(7); 2901-2907. [PubMed: 7911787].

7. Product Monograph: Dukoral: Product Monograph: Dukoral vaccine information [http://healthsensetravelclinic.ca/wp-content/uploads/dukoral.pdf]

8. Ravichandran et al., 2006: Ravichandran M, Ali SA, Rashid NH, Kurunathan S, Yean CY, Ting LC, Bakar AS, Lalitha P, Zainuddin ZF. Construction and evaluation of a O139 Vibrio cholerae vaccine candidate based on a hemA gene mutation. Vaccine. 2006; 24(18); 3750-3761. [PubMed: 16102875].

9. Sharma et al., 2008: Sharma MK, Singh NK, Jani D, Sisodia R, Thungapathra M, Gautam JK, Meena LS, Singh Y, Ghosh A, Tyagi AK, Sharma AK. Expression of toxin co-regulated pilus subunit A (TCPA) of Vibrio cholerae and its immunogenic epitopes fused to cholera toxin B subunit in transgenic tomato (Solanum lycopersicum). Plant cell reports. 2008; 27(2); 307-318. [PubMed: 17962948].

10. Vishwakarma et al., 2015: Vishwakarma V, Sahoo SS, Das S, Ray S, Hardt WD, Suar M. Cholera toxin-B (ctxB) antigen expressing Salmonella Typhimurium polyvalent vaccine exerts protective immune response against Vibrio cholerae infection. Vaccine. 2015; 33(15); 1880-1889. [PubMed: 25701672].

11. WHO: Cholera: WHO: Cholera [http://www.who.int/topics/cholera/about/en/index.html]

12. Xu et al., 2009: Xu G, Wang S, Zhuang L, Hackett A, Gu L, Zhang L, Zhang C, Wang H, Huang Z, Lu S. Intramuscular delivery of a cholera DNA vaccine primes both systemic and mucosal protective antibody responses against cholera. Vaccine. 2009; 27(29); 3821-3830. [PubMed: 19443090].

13. Yu et al., 2005: Yu F, Qi G, Liu Y, Gao S, Kan B. Construction and characterization of a thyA mutant derived from cholera vaccine candidate IEM101. Molecular biotechnology. 2005; 29(3); 191-196. [PubMed: 15767696].