supplemental figure 1a. sds-page analysis showing omps expressed by v. tubiashii strain 19105. from...
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Supplemental Figure 1a. SDS-PAGE analysis showing OMPs expressed by V. tubiashii strain 19105. From left to right, Lane 1, cells grown in the presence of 3% NaCl compared to those grown with 1% NaCl in Lane 2; Lane 3 cells grown in the presence of 0.3% bile salts compared to those grown without bile salts Lane 4; Lane 5 cells grown in the presence of 2% maltose compared to those grown without maltose Lane 6; Lane 7 represents the molecular markers, sizes are reported in kDa. Note the sequenced and uniquely expressed OMPs at each environmental condition are identified with an arrow.
50 kDa
70 kDa
45 kDa
40 kDa
90 kDa
25 kDa
20 kDa
15 kDa
65 kDa
35 kDa
14.4
20.1
30.0
43.0
67.0
94.0
Bile salts
0.3% 0%
NaCl
3% 1%
Maltose
2% 0% MW
1a.
14.4
20.1
30.0
43.0
67.0
94.0
Temperature
37° 30°
pH
8.5 7 6 MW
50 kDa
70 kDa
45 kDa
40 kDa
90 kDa
25 kDa
20 kDa
15 kDa
65 kDa
35 kDa
Supplemental Figure 1b. SDS-PAGE analysis showing OMPs expressed by V. tubiashii strain 19105. From left to right, Lane 1-2, cells grown at different temperatures; and Lane 3-5, cells grown at various pH conditions; Lane 6 represents the molecular markers, sizes are reported in kDa. Note the sequenced and uniquely expressed OMPs at each environmental condition are identified with an arrow.
1b.
Supplemental Figure 2a. SDS-PAGE analysis showing OMPs expressed by V. tubiashii strain 19109. From left to right, Lane 1, cells grown in the presence of 3% NaCl compared to those grown with 1% NaCl Lane 2; Lane 3, cells grown in the presence of 0.3% bile salts compared to those grown without bile salts Lane 4; Lane 5, cells grown in the presence of 2% maltose compared to those grown without maltose, Lane 6; Lane 7 represents the molecular markers, sizes are reported in kDa. Note the sequenced and uniquely expressed OMPs at each environmental condition are identified with an arrow.
50 kDa
70 kDa
45 kDa40 kDa
90 kDa
25 kDa
20 kDa
15 kDa
65 kDa
35 kDa
14.4
20.1
30.0
43.0
67.0
94.0
NaCl3% 1%
Bile salts0.3% 0%
Maltose2% 0% MW
2a.
14.4
20.1
30.0
43.0
67.0
94.0
Temperature37° 30°43°
pH8.5 7 6 MW
50 kDa
70 kDa
45 kDa40 kDa
90 kDa
25 kDa
20 kDa
15 kDa
65 kDa
35 kDa
Supplemental Figure 2b. SDS-PAGE analysis showing OMPs expressed by V. tubiashii strain 19109. From left to right, Lane 1-3, cells grown at various temperatures; and Lane 4-6, cells grown at various pH conditions; Lane 7 represents the molecular markers, sizes are reported in kDa. Note the sequenced and uniquely expressed OMPs at each environmental condition are identified with an arrow.
2b.
a.
Vv4965-T1
CTCTAGAAGTTGGTGGCCGTGCTGAAGCTCGCCTATCTCTGAAAGATGGTAAGGCACAAG
Vc395
CTCTAGAAGTTGGTGGCCGTGCTGAAGCTCGCCTATCTCTGAAAGATGGTAAGGCACAAG
Vt109
CTCTAGAGATCGGCGGTCGTGCTGAAGCTCGTATGTCTCTAAAAGATGGTAAAGCTGAAG
Vt105
CTATCGAAATGGGTGGTCGCGCGGAAGCACGTCTAGCTCTTAAAGATGGNAAGGCAGAAN
** * ** * ** ** ** ** ***** ** * **** ******** ** ** **
Vv4965-T1 ACAACTCTCGCGTACGTCTAAACTTCTTGGGTAAAGCAGAAATCAATGACAGCCTATACG
Vc0395 ACAACTCTCGCGTACGTCTAAACTTCTTGGGTAAAGCAGAAATCAATGACAGCCTATACG
Vt109 ACAAAACACGTGTACGTCTAAACTTCCTAGGTAAGACTCAAATCACTGATAGCCTATATG
Vt105 ACAAATCTCGCGTACGTCTAAACTTCCTAGGTACAGCTCAAATCACTGACAGCCTATACG
**** * ** *************** * **** * ****** *** ******** *
Vv4965-T1 GTGTTGGTTTCTACGAAGGCGAGTTTACTACTAATGATCA------------AGGTAAAA
Vc395 GTGTTGGTTTCTACGAAGGCGAGTTTACTACTAATGATCA------------AGGTAAAA
Vt109 GTCTAGGTTTCTACGAAGCTGAGTACACTACAGCAGACAACACTGTAACTGTAGACGGCA
Vt105 GTGTGGGTTTCTACGAAGGTGAGTTCACTACTAATGACAATGGCGGCGCTACAGATGGCA
** * ************* **** ***** ** * ** *
Vv4965-T1 ACG-CGTCTAACA---ACAGCCTANACAACCGTTATACCTACGCTGGTATCGGTGGCACT
Vc395 ACG-CGTCTAACA---ACAGCCTAGACAACCGTTATACCTACGCTGGTATCGGTGGCACT
Vt109 AAA-CAACTGACGGTGATAACATTGAGCACCGTTACACTTACGCTGGTATTGGTGGTGAC
Vt105 ACAACAGCTCATC----TGATCTAACTAACCGTTACCTATACGGTGGTATCGGCGGAGCT
* * ** * * ******* **** ****** ** **
Vv4965-T1 TACGGTGAAGTGACTTACGGTAAAAACGATGGCGCATTGGGCGTAATCACTGACTTCACC
Vc395 TACGGTGAAGTGACTTACGGTAAAAACGATGGCGCATTGGGCGTAATCACTGACTTCACC
Vt109 TTTGGTGAAGTAACTTACGGTAAAAACGATGGTGCACTANGCGTTATCACTGACTTTACT
Vt105 TTCGGTGAAGTAACTTACGGTAAGAACGACGGTGCGCTANGTGTTATCACTGATTTCACT
* ******** *********** ***** ** ** * * ** ******** ** **
Supplemental Figure 3. Clustal X alignment of nucleotide sequence of ompU gene from V. tubiashii strains 19105 and 19109, V. vulnificus 4965-T1 and V. cholerae 395, respectively. ompU gene sequences were obtained in this study. Indicated by underlying are the PCR primer sequences used in the study. a. Forward sequence, b. Reverse sequence. The symbol “*” represents conserved nucleotides among the sequences.
Vv4965-T1 NCNNTGGACTGACC-CAGTTCGTC-TTCTGATGCTACTTTACCAA----------CTTTG
Vc395 GCNNNGNANTTANG-CAATACNTNGTTCTGATGCTACTTTACCAAA---------CTTTG
Vt109 GCNNNGAACTGAANATAGCTCGTC-TTCTGCACCAGCTTTACCTACAGTACCAGTTGCTG
Vt105 NCCNTNGACNNAAA-AAGCTCGTC-TTCTGCATCTGCTTTAGAGATG----TAAGATTTG
* * * * * * ***** * ***** * **
Vv4965-T1 TCTGA--ATCT------AGCAGATTGAACTGGTAAGAGATGTAAGAGCGNNAGTTTGGCT
Vc395 GCTGA--ATCT------AGCAGATTGAACTGGTAAGAGATGTAAGAGCGNAAGTTTGGCT
Vt109 TCTTATCATCTTTACTAAGTAGGTTGAAGTTGTAAGAGATGTAAGTACGGAAGTTTGGCT
Vt105 TCGTC--GTCT------AGCAGGTTAAAGTTGTACGATACGTAGNCACGGAAGTTAGGCT
* *** ** ** ** ** * *** ** * *** ** **** ****
Vv4965-T1 TGAAGTAGTAAGTTGCGTCGATAGCAAAATTATCTGCTGAAGT--------T-TCTTTCG
Vc395 TGAAGTAGTAAGTTGCGTCGATAGCAAAATTATCTGCTGAAGT--------TCTCTTTCG
Vt109 TGAAGTAGTATGTTGCATCGATCGCTACATTGTCAGCTGCATCATACGTAGTACCTGAAG
Vt105 TAAAGTAGTATGTTGCGTCGATAGCGATGTTATCTGCAGCATCTTCTTTAGTTGAACCTG
* ******** ***** ***** ** * ** ** ** * * * *
Vv4965-T1 CTGTTTCTGCA---TTGTTGTATGTCGCAGTAAACGCAGCTTGACCTAGTTTGTAACCAG
Vc395 CTGTTTCTGCA---TTGTTGTATGTCGCAGTAAACGCAGCTTGACCTAGTTTGTANCCAG
Vt109 TTTTCTCTTCTAGCTTATTGTATGTTGTAGTGAATGCTACTTTTTCTAGAGTGTAAGCTG
Vt105 TTTTGTTTTCTAGGTAGTTGTAAGACGTTGTGAATACAGTCTGGCCCATAGTGTAAGCTG
* * * * * ***** * * ** ** * * * * ***** * *
Vv4965-T1 CAGCTAGCTCGTAGCCAGTGTAGTCAACGT-----CTTTCGC---TAGCTC-------AC
Vc395 CANCTANCTCGTAGCCAGTGTAGTCAACGT-----CTTNCGC---TAGCTC-------AC
Vt109 CTGCGAATTCGTAACCAGTTTTGTCTGTTTT---GCTTGAGC---CGTCTTTGTCTTTAC
Vt105 CTGCTAGTTCGTAACCTGTCTTATCAGTTGTTGAGCTAGAACCTTTCACTTGGTCTTCGC
* * * ***** ** ** * ** ** * ** *
b.
Supplemental Figure 3. Clustal X alignment of nucleotide sequence of ompU gene from V. tubiashii strains 19105 and 19109, V. vulnificus 4965-T1 and V. cholerae 395, respectively. ompU gene sequences were obtained in this study. Indicated by underlying are the PCR primer sequences used in the study. a. Forward sequence, b. Reverse sequence. The symbol “*” represents conserved nucleotides among the sequences.
Vt109 NCNNNNTCCNNNNCTTACCCAGCCTGACTCAAGCCATCTCAACCCTACGTAAAATGCTGA
Vt105 ----------NCNNNTTCAAAACCTGACTCAGGCCATCTCATCCCTACGTAAAATGCTGA
Vf33809 ---------------TGGTGGAATGCACACCGGCGGACTCGGCCCCAAAGAAGTCATCGC
* ** * ** * *** *** * ** *
Vt109 AAGACTCGACCAAGTCGCC--TCAGTTTGTGAAA-ACGGTTCCAAAACGTGGTTATCAGT
Vt105 AAGACTCGACTAAATCCCCCATCAATCCGTGGAAGACGGTTCCAAAACGTGGTTATCAGT
Vf33809 TACCGGCGGTCAAAAC--------------------------------------------
* ** **
Vt109 TGATCGCGACCGTCGAAAGCGTTCAGCTCGATGCAAGTAAAGATCCTGATGCCATCGAGC
Vt105 TGATCGCGACCGTCGAAACCCTGGAGCTCGATGCAAGTAAAGATCCTGATGCCATCGAGC
Vf33809 ------------------------------------------------------------
Vt109 AGTCAGATGAGTCTTACTCAAGTGCCGTATCCTCTTCAGAACTTGTCAGTCAAGCTGCTG
Vt105 AATCATATGAGTCTTACTCAAGTGCCGTATCCTCTTCACAACTTGTCAGTCAAGCTGCTG
Vf33809 ------------------------------------------------------------
Vt109 TTTCTGAAGCAGTTATTACTCAGACGTCGCCTTCTGTAGACTCGACACCTGCGGCACCAA
Vt105 TTTCTGAATCAGTTATTACTCAGACGTCGCCTTCTGTAGACTCGACACCTGCAGCACCAA
Vf33809 ------------------------------------------------------------
Vt109 AACCACGAATGGATCTGTTCACTAAATTGATTCTGTTGGTTGCCGTCATACTGCCAT---
Vt105 AACCACGAATGGATCTGTTCACTAAATTGATTCTGTTGGTTGCCGTCATACTGCCATCNN
Vf33809 ------------------------------------------------------------
a.
Supplemental Figure 4. Clustal X alignment of nucleotide sequence of toxR gene from V. tubiashii strains 19105 and 19109 compared to that of V. cholerae 395 and V. fluvialis strain ATCC 33809. toxR gene sequences were obtained in this study and gene sequence for V. fluvialis strain ATCC 33809 was obtained from the NCBI accession number AF170885. a. Forward sequence, b. Reverse sequence. The symbol “*” represents conserved nucleotides among the sequences.
Vt109 TGAACAGATCCATTCGTGGTTTTGGTGCCGCAGGTGTCGAGT-CTACAGAAGGCGACGTC
Vt105 TAAAGGATAACCGTAAGGATGTCGGAGTTCCAATTATTACGCACTCCAGCAGTCGATGTG
Vf33809 TGGACGACTCCAGCCTGACTCAAGCCATCTCAACC--------CTACGTAAAATGCTGAA
* * * * * ** ** * * * *
Vt109 TGAGTAATAACTGCTTCAGAAACA-------GCAGCTTG-ACTGACAAGTTCTGAAGAGG
Vt105 TGAGCCAAGAGGACTCCATGAACACTTAGNCGCGGTTTCTGCGGACCGATTCCGTGAAGG
Vf33809 AGACT--CGACCAAGTC---GCCT--------CAGTTTGTGAAAAC-GGTTCCAAAACGT
** * * * * ** * *** *
Vt109 AT------------ACGGTACTTGAGTAAGACTCATCTGACTGCTCGATG--GCATCAGG
Vt105 ATGGATAGGGCCGGACGATAATTGTGCAAAATTAACTTGACTGCTCGATTATGCATCAGG
Vf33809 GGTTATCAGTT--GATCGCGACCGTCGAAAGCGTTCAGCTCGATGCAAGTAAAGATCCTG
* * ** * * * ** *
Vt109 ATCT-TTACTT--GCATCGAGC-TGAACGCTT-TCGACGGTCGCGATC--AACTGATAAC
Vt105 ATCTATTACCTATGCATCGAGCCTGAACGCTTATTAACGCTCGCTACCCNGAGTGATAAC
Vf33809 ATGC--CATCGA-GCAGTCAGA-TGAGTCTTACTCAAGTGCCGTATCC--TCTTCAGAAC
** * *** ** *** * * * ** * * * ***
Vt109 CAC---GTTTTGGAACCG--TTT-----TCACAAACTGAGGCGACTTGGTCGAGT-----
Vt105 CACAGTGGTTGGTAACCGGATTTGNCGATCATAAATGGGTGCGACTTGGTGGTCTAGCTC
Vf33809 TTGTCAGTCAAGCTGCTG---TT-----TCTGAAGCAGTTATTACTCAGACGTCG-----
* * * * ** ** ** * *** * *
Vt109 -CTTTCAGCATTTTACGTA------GGGTTGAG-ATGGCTTGAGTCAGGCTGGA--GTCG
Vt105 GCTCTCAAACTTTTACAAACGTTAGGGGTTGAACATGGCTTGACTCAGGCTGCAACGTCT
Vf33809 -CCTTCTGTAGACTCGACACCTGCGGCACCAAA---ACCACGAATGGATCTGTT----CA
* ** * * * ** *** *
Vt109 TCCACCT-CAAACCCCTGAG---AATACGTCTCCCNCNCTCTTNCAATCTCACAGTTCCA
Vt105 TCCACCTTCAAACCCCTGAACCAAATACCTGTACTNCNCTCTTNCGTTTCCATAGTCACA
Vf33809 CTAAATT--GATTCTGTTGGT--TGCCGTCATACTGCCTATTTGTGTCATTTTGTTTACG
* * * * * * ** * *
b.
Supplemental Figure 4. Clustal X alignment of nucleotide sequence of toxR gene from V. tubiashii strains 19105 and 19109 compared to that of V. cholerae 395 and V. fluvialis strain ATCC 33809. toxR gene sequences were obtained in this study and gene sequence for V. fluvialis strain ATCC 33809 was obtained from the NCBI accession number AF170885. a. Forward sequence, b. Reverse sequence. The symbol “*” represents conserved nucleotides among the sequences.