Browse > Article
http://dx.doi.org/10.4014/jmb.1111.11016

Identification of Virulence Factors in Vibrio vulnificus by Comparative Transcriptomic Analyses between Clinical and Environmental Isolates Using cDNA Microarray  

Kim, In-Hwang (Department of Life Science and Interdisciplinary Program of Integrated Biotechnology, Sogang University)
Kim, Byung-Soo (Department of Applied Statistics, Yonsei University)
Lee, Kyung-Shin (Department of Life Science and Interdisciplinary Program of Integrated Biotechnology, Sogang University)
Kim, Ik-Joong (Department of Life Science and Interdisciplinary Program of Integrated Biotechnology, Sogang University)
Son, Jee-Soo (Department of Life Science and Interdisciplinary Program of Integrated Biotechnology, Sogang University)
Kim, Kun-Soo (Department of Life Science and Interdisciplinary Program of Integrated Biotechnology, Sogang University)
Publication Information
Journal of Microbiology and Biotechnology / v.21, no.12, 2011 , pp. 1228-1235 More about this Journal
Abstract
We compared the gene expression among four clinical and five environmental V. vulnificus isolates, using a cDNA microarray containing 131 genes possibly associated with pathogenicity, transport, signal transduction, and gene regulations in the pathogen. cDNAs from total RNAs of these isolates were hybridized into the cDNA microarray using the cDNA of the wild-type strain MO6-24/O as a reference. We focused on selecting differentially expressed (DE) genes between clinical and environmental isolates using a modified t-statistic. We could detect two statistically significant DE genes between virulent isolates and less-virulent isolates with a marginal statistical significance (p-value of 0.008). These were genes putatively encoding pilin and adenlyate cylase. Real time-PCR confirmed that these two selected genes transcribed in significantly higher levels in virulent isolates than in less-virulent isolates. Mutants with lesions in the gene encoding pilin showed significantly higher $LD_{50}$ values than that of wild type.
Keywords
Vibrio vulnificus; virulence factors; cDNA microarray; pilin;
Citations & Related Records
Times Cited By KSCI : 1  (Citation Analysis)
Times Cited By Web Of Science : 0  (Related Records In Web of Science)
연도 인용수 순위
1 Dudoit, S., J. P. Shaffer, and J. C. Boldrick. 2003. Multiple hypothesis testing in microarray experiments. Stat. Sci. 18: 71-103.   DOI   ScienceOn
2 Fujita, A., J. R. Sato, L. O. Rodrigues, C. E. Ferreira, and M. C. Sogayar. 2006. Evaluating different methods of microarray data normalization. BMC Bioinformatics 7: 469.   DOI
3 Glickmann, E., L. Gardan, S. Jacquet, S. Hussain, M. Elasri, A. Petit, and Y. Dessaux. 1998. Auxin production is a common feature of most pathovars of Pseudomonas syringae. Mol. Plant Microbe Interact. 11: 156-162.   DOI   ScienceOn
4 Jeong, H. S., H. H. Lee, K.-H. Lee, S.-J. Park, and S. H. Choi. 2003. SmcR and cyclic AMP receptor protein coactivate Vibrio vulnificus vvpE encoding elastase through the RpoS-dependent promoter in a synergistic manner. J. Biol. Chem. 278: 45072-45081.   DOI
5 Johnson, D. E., F. M. Calia, D. M. Musher, and A. Goree. 1984. Resistance of Vibrio vulnificus to serum bactericidal and opsonizing factors: Relation to virulence in suckling mice and humans. J. Infect. Dis. 150: 413-418.   DOI   ScienceOn
6 Kim, I. H., J.-I. Shim, K.-E. Lee, W. Hwang, I.-J. Kim, S.-H. Choi, and K.-S. Kim. 2008. Nonribosomal peptide synthase is responsible for the biosynthesis of siderophore in Vibrio vulnificus MO6-24/O. J. Microbiol. Biotechnol. 18: 35-42.
7 Rosche, T. M., Y. Yano, and J. D. Oliver. 2005. A rapid and simple PCR analysis indicates there are two subgroups of Vibrio vulnificus which correlate with clinical or environmental isolation. Microbiol. Immunol. 49: 381-389.
8 Schena, M., R. A. Heller, K. Theriault, T. P. Konrad, E. Lachenmeier, and R. W. Davis. 1999. Microarrays: Biotechnology's discovery platform for functional genomics. Trends Biotechnol. 17: 217-218.   DOI   ScienceOn
9 Smyth, G. K. 2004. Linear models and empirical Bayes methods for assessing differential expression in microarray experiments. Stat. Appl. Genet. Mol. Biol. 3: Article 3.
10 Stelma Jr., G. N., A. L. Reyes, J. T. Peeler, C. H. Johnson, and P. L. Spaulding. 1992. Virulence characteristics of clinical and environmental isolates of Vibrio vulnificus. Appl. Environ. Microbiol. 58: 2776-2782.
11 Strom, M. S. and R. N. Paranjpye. 2000. Epidemiology and pathogenesis of Vibrio vulnificus. Microbes Infect. 2: 177-188.   DOI   ScienceOn
12 Morris, J. G. and R. E. Black. 1985. Cholera and other vibrioses in the United States. N. Engl. J. Med. 312: 343-350.   DOI   ScienceOn
13 Blake, P. A., M. H. Merson, R. E. Weaver, D. G. Hollis, and P. C. Heublein. 1979. Disease caused by a marine Vibrio. Clinical characteristics and epidemiology. N. Engl. J. Med. 300: 1-5.   DOI   ScienceOn
14 Churchill, G. A. 2002. Fundamentals of experimental design for cDNA microarrays. Nature Genet. (Supplement) 32: 490-495.   DOI   ScienceOn
15 Kreger, A., L. DeChatelet, and P. Shirley. 1981. Interaction of Vibrio vulnificus with human polymorphonuclear leukocytes: Association of virulence with resistance to phagocytosis. J. Infect. Dis. 144: 244-248.   DOI   ScienceOn
16 Lee, S. E., P. Y. Ryu, S. Y. Kim, Y. R. Kim, J. T. Koh, O. J. Kim, et al. 2004. Production of Vibrio vulnificus hemolysin in vivo and its pathogenic significance. Biochem. Biophys. Res. Commun. 324: 86-91.   DOI   ScienceOn
17 Milton, D. L. 1996. Flagellin A is essential for the virulence of Vibrio anguillarum. J. Bacteriol. 178: 1310-1319.
18 Miyoshi, S. 2006. Vibrio vulnificus infection and metalloprotease. J. Dermatol. 33: 589-595.   DOI   ScienceOn
19 Panicker, G., D. R. Call, M. J. Krug, and A. K. Bej. 2004. Detection of pathogenic Vibrio spp. in shellfish by using multiplex PCR and DNA microarrays. Appl. Environ. Microbiol. 70: 7436-7444.   DOI   ScienceOn
20 Park, D.-K., K.-E. Lee, C.-H. Baek, I. H. Kim, J.-H. Kwon, W. K. Lee, et al. 2006. Cyclo(Phe-Pro) modulates the expression of ompU in Vibrio spp. J. Bacteriol. 188: 2214-2221.   DOI   ScienceOn
21 Webster, A. C. and C. M. Litwin. 2000. Cloning and characterization of vuuA, a gene encoding the Vibrio vulnificus ferric vulnibactin receptor. Infect. Immun. 68: 526-534.   DOI   ScienceOn
22 Taylor, L. A. and R. E. Rose. 1988. A correction in the nucleotide sequence of the Tn903 kanamycin resistance determinant in pUC4K. Nucleic Acids Res. 16: 358.   DOI   ScienceOn
23 Tison, D. L. and M. T. Kelly. 1986 Virulence of Vibrio vulnifiucs strains from marine environments. Appl. Environ. Microbiol. 51: 1004-1006.
24 Troyanskaya, O., M. Cantor, G. Sherlock, P. Brown, T. Hastie, R. Tibshirani, et al. 2001. Missing value estimation methods for DNA microarrays. Bioinformatics 17: 520-525.   DOI   ScienceOn
25 Wolfgan, M. C., V. T. Lee, M. E. Gilmore, and S. Lory. 2003. Coordinate regulation of bacterial virulence genes by a novel adenylate cyclase-dependent signal pathway. Dev. Cell 4: 253-263.   DOI   ScienceOn
26 Yoshida, S.-I., M. Ogawa, and Y. Mizuguchi. 1985. Relation of capsular materials and colony opacity to virulence of Vibrio vulnificus. Infect. Immun. 47: 446-451.
27 Wright, A. C., J. G. Morris Jr., D. R. Maneval Jr., K. Richardson, and J. B. Backer. 1985. Cloning of the cytotoxinhemolysin gene of Vibrio vulnificus. Infect. Immun. 50: 922-924.
28 Yamashiro, T., N. Nakasone, and M. Iwanaga. 1993. Purification and characterization of pili of a Vibrio cholera Non-O1 strain. Infect. Immun. 61: 5398-5400.
29 Yang, Y. H., S. Dudoit, P. Luu, D. M. Lin, V. Peng, J. Ngai, and T. P. Speed. 2002. Normalization for cDNA microarray data: A robust composite method addressing single and multiple slide systematic variation. Nucleic Acids Res. 30: e15.   DOI
30 Yuan, J. S., A. Reed, F. Chen, and C. N. Stewart Jr. 2006. Statistical analysis of real-time PCR data. BMC Bioinformatics 7: 85.   DOI