從創傷弧菌Vibrio vulnificus CKM-1選殖(一)藍色螢光蛋白基因bfpvv，與(二)胞外酵素脂解酶的基因lipA及其下游基因lipB ；並分別加以定序及進行特性分析。（一）從創傷弧菌Vibrio vulnificus CKM-1將藍色螢光蛋白基因bfpvv選殖並定序。當使用紫外線(UV, 245 ~ 366 nm)照射時，轉形體表現藍色螢光。經核苷酸序列分析，推論一段長度含717 bp的開放讀框架(open reading frame, ORF)可編碼一段長度含239個胺基酸的胜肽鏈，分子質量估算約25.8 kDa。根據bfpvv基因的核苷酸序列及其推測的胺基酸序列，顯示其與來自各種生物體的短鏈去氫酶/還原酶家族(short chain dehydrogenase/reductase family, SDR family)蛋白質有顯著的同源性。短鏈去氫酶/還原酶上有一些重要的功能性殘基被嚴格的保留在Vibrio vulnificus CKM-1的藍色螢光蛋白BFPVV上，如活化位的Tyr145，催化位的Lys149，及特定的N端序列GlyXXXGlyXGly等。分別將這些特定的胺基酸殘基改變：Tyr145改為Phe，Lys149改為Ile，而Gly9改為Val；發現G9V突變株不產生藍色螢光，而與野生株相對比較，Y145F突變株產生126％的螢光強度，K149I突變株則產生68.5％的螢光強度。
（二）從Vibrio vulnificus CKM-1將胞外酵素脂解酶的基因lipA及其下游基因lipB選殖並加以定序。經核苷酸序列及胺基酸序列比對，我們認為LipA蛋白質是細菌性脂解酶家族I.1的組成員之一，而LipB蛋白質則是LipA脂解酶的活化子。在Escherichia coli細胞轉型體中，LipA與LipB必須共存時才具有脂解酶的活性。對其基質作探討，LipA分別利用對硝基酚酯(p-nitrophenyl esters)及三酸甘油酯(triacylglycerols) 為基質，進行水解反應試驗，發現當基質為對硝基酚酯時，其水解反應之最適當醯鏈長為C14，而當受質為三酸甘油酯時，其水解反應之最適當鏈長為C10到C12。

The blue fluorescent protein gene (bfpvv) and extracellular lipase gene (lipA) with its downstream gene (lipB) from Vibrio vulnificus CKM-1 were cloned, sequenced, and characterized respectively.
1. The blue fluorescent protein (BFPVV) gene, bfpvv, from Vibrio vulnificus CKM-1 was cloned and sequenced. Expression of the bfpvv gene in Escherichia coli XL1B could be visualized through the illumination with UV source. Nucleotide sequence analysis predicted a single open reading frame of 717 bp encoding a 239-amino-acid polypeptide with a calculated molecular mass of 25.8 kDa. The BFPVV protein has been produced in E. coli XL1B or E. coli BL21. The nucleotide sequence of bfpvv gene and its deduced amino acid sequence showed significant homology to those of the short-chain dehydrogenase / reductase (SDR) family proteins from various organisms. Some functionally important residues in SDR were strictly conserved in BFPVV, such as an active site-Tyr145, a catalytic site-Lys149 and a common GlyXXXGlyXGly pattern in the N-terminal part of the molecule. By changing three amino acid residues, Tyr145, Lys149, and Gly9 to Phe, Ile, and Val, respectively, it was found that G9V mutant did not generate blue fluorescence, while the mutants Y145F and K149I have 126% and 68.5% fluorescence compared with the wild-type BFPVV.
2. The gene (lipA) encoding the extracellular lipase and its downstream gene (lipB) from Vibrio vulnificus CKM-1 were cloned and sequenced. Nucleotide sequence analysis and alignments of amino acid sequences suggest that LipA is a member of family I.1 lipase and that LipB is a lipase activator of LipA. The active LipA was produced in recombinant Escherichia coli cells only in the presence of the lipB. In the hydrolysis of p-nitrophenyl esters using the reactivated LipA, optimum chain lengths for the acyl moiety on the substrate were C12 to C14.

References:
Alm, R. A., Stroeher, U. H. and Manning, P. A. Extracellular proteins of Vibrio cholerae: nucleotide sequence of the structural gene (hlyA) for the haemolysin of the haemolytic E1 Tor strain 017 and characterization of the hlyA mutation in the non-haemolytic classical strain 569B. Mol. Microbiol. 2 (4), 481-488. (1988)
Arpigny, J. L. and Jaeger, K.-E. Bacterial lipolytic enzymes: classification and properties. Biochem. J. 343, 177-183. (1999)
Baker, M. Protochlorophyllide reductase is homologous to human carbonyl reductase and pig 20β-hydroxysteroid dehydrogenase. Biochem. J. 300, 605-607. (1994)
Baldwin, T. O., Devine, J. H., Heckel, R. C., Lin, J. W. and Shadel, G. S. The complete nucleotide sequence of the lux regulon of Vibrio fischeri and the luxABN region of Photobacterium leiognathi and the mechanism of control of bacterial bioluminescence. J. Biolumin. Chemilumin. 4, 326-341. (1989)
Baldwin, T. O., Treat, M. L. and Daubner, S. C. Cloning and expression of the luxY gene from Vibrio fischeri strain Y-1 in Escherichia coli and complete amino acid sequence of the yellow fluorescent protein. Biochem. 29, 5509-5515. (1990)
Baron, S. F., Franklund, C. V. and Hylemon, P. B. Cloning, sequencing, and expression of the gene coding for bile acid 7α-hydroxysteroid dehydrogenase from eubacterium sp. strain VPI 12708. J. Bacteriol. 173, 4558-4569. (1991)
Bassler, B. L., Greenberg, E. P. and Stevens, A. M. Cross-species induction of luminescence in the quorum-sensing bacterium Vibrio harveyi. J. Bacteriol. 179, 4043-4045. (1997)
Baumann, P., and Baumann, L., Woolkalis, M. and Bang, S. Evolutionary relationships in Vibrio and Photobacterium. A basis for a natural classification. Ann. Rev. Microbiol. 37, 369-398. (1983)
Beggs, J. D. Transformation of yeast by a replicating hybrid plasmid. Nature 275, 104-109. (1978)
Belas, R., Mileham, A., Cohn, D., Hilmen, M., Simon, M. and Silverman, M. Bacterial bioluminescence: isolation and expression of the luciferase genes from Vibrio harveyi. Science 218, 791-793. (1982)
Bisharat, N., Agmon, V., Finkelstein, R., Raz, R., Ben Dror, G., Lerner, L., Soboh, S., Colodner, R., Cameron, D. N., Wykstra, D. L., Swerdlow, D. L. and Farmer, III. J.J. Clinical, epidemiological, and microbiological features of Vibrio vulnificus biogroup 3 causing outbreaks of wound infection and bacteraemia in Israel. Israel Vibrio Study Group. Lancet. 354, 1421-1424. (1999)
Blake, P. A., Merson, M. H., Weaver, R. E., Hollis, D. G., and Heublein, P. C. Disease caused by a marine Vibrio. Clinical characteristics and epidemiology. N. Engl. J. Med. 300, 1-5. (1979)
Bolivar, F., Rodriguez, R. L., Greene, P. J., Betlach, M. C., Heyneker, H. L. and Boyer, H. W. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2, 95-113. (1977)
Borenstein, M. and Kerdel, F. Infections with Vibrio vulnificus. Dermatol Clin. 21(2), 245-8. (2003)
Brennt, C. E., Wright, A. C., Dutta, S. K. and Morris, Jr. J. G. Growth of Vibrio vulnificus in serum from alcoholics: association with high transferrin iron saturation. J. Infect. Dis. 164, 1030-1032. (1991)
Breton, R., Housset, D., Mazza, C. and Fontecilla-Camps, J. C. The structure of a complex of human 17beta-hydroxysteroid dehydrogenase with estradiol and NADP+ identifies two principal targets for the design of inhibitors. Structure 4, 905-915. (1996)
Bullen, J. J., Spalding, P. B., Ward, C. G., and Gutteridge, J. M. Hemochromatosis, iron and septicemia caused by Vibrio vulnificus. Arch. Intern. Med. 151, 1606-1609. (1991)
Bush, C. A., Patel,P., Gunawardena, S., Powell, J., Joseph, A., Johnson, J. A., and Morris, J. G. Classification of Vibrio vulnificus strains by the carbohydrate composition of their capsular polysaccharides. Anal. Biochem. 250, 186-195. (1997)
Chalfie, M. Green fluorescent protein. Photochem. Photobiol. 62, 651-656. (1955)
Chalfie, M., Tu, Y., Euskirchen, G., Ward, W. W. and Prasher, D. C. Green fluorescent protein as a marker for gene expression. Science 263, 802-805. (1994)
Chan, T. Y., Chow, D. P., Ng, K. C., Pan, K. W. and McBride, G. A. Vibrio vulnificus septicemia in a patient with liver cirrhosis. Southeast Asian J. Trop. Med. Public Health. 25, 215-216. (1994)
Chang, M. C., Chang, J. C. and Chen, J. P. Cloning and nucleotide sequence of an extracellular α-amylase gene from Aeromonas hydrophila MCC-1. J. Gen. Microbiol. 139, 3215-3223. (1993)
Chang, T. M., Chuang, Y. C., Su, J. H. and Chang, M. C. Cloning and sequence analysis of a novel hemolysin gene (vllY) from Vibrio vulnificus. Appl. Environ. Microbiol. 63, 3851- 3857. (1997)
Chavagnat, F., Duez, C., Guinand, M., Potin, P., Barbeyron, T., Henrissat, B., Wallach, J. and Ghuysen, J. M. Cloning, sequencing and overexpression in Escherichia coli of the alginatelyase-encoding aly gene of Pseudomonas alginovora: identification of three classes of alginate lyases. Biochem. J. 319, 575-583. (1996)
Chen, Z., Jiang, J. C., Lin, Z. G., Lee, W. R., Baker, M. E. and Chang, S. H. Site-specific mutagenesis of Drosophila alcohol dehydrogenase: Evidence for involvement of tyrosine-152 and Lysine-156 in catalysis. Biochem. 32, 3342-3346. (1993)
Chen, Y. C., Chang, M. C., Chuang, Y. C. and Jeang, C. L. Characterization and virulence of hemolysin III from Vibrio vulnificus. Curr. Microbiol. 49, 175-179. (2004)
Chen, Z., Lu, L., Shirley, M., Lee, W. R. and Chang, S. H. Site-directed mutagenesis of Glysine-14 and two“critical”Cysteinyl residues in Drosophila alcohol dehydrogenase. Biochem. 29, 1112-1118. (1990)
Chuang, Y. C., Chang, T. M. and Chang, M. C. Cloning and characterization of the gene (empV) encoding extracellular metalloprotease from Vibrio vulnificus. Gene 189, 163-168. (1997)
Chuang, Y. C., Chiou, S. F., Su, J. H., Wu, M. L. and Chang, M. C. Molecular analysis and expression of the extracellular lipase of Aeromonas hydrophila MCC-2. Microbiol 143, 803-812. (1997)
Chuang, Y. C., Sheu, H. M., Ko, W. C., Chang, T. M., Chang, M. C. and Huang, K. Y. Mouse skin damage caused by a recombinant extracellular metalloprotease from Vibrio vulnificus and by V. vulnificus infection. J. Formos Med. Assoc. 96(9), 677-684. (1997)
Cohn, D. H., Mileham, A. J., Simon, M. I., Nealson, K. H., Rausch, S. K., Bonam, D. and Baldwin, T. O. Nucleotide sequence of the luxA gene of Vibrio harveyi and the complete amino acid sequence of the α subunit of bacterial luciferase. J. Biol. Chem. 260, 6139-6146. (1985)
Coleman, J. P., White, W. B., Lijewski, M. and Hylemon, P. B. Nucleotide sequence and regulation of a gene involved in bile acid 7-dehydroxylation by Eubacterium sp. Strain VPI 12708. J. Bacteriol. 170, 2070-2077. (1988)
Daubner, S. C., Astorga, A. M., Leisman, G. B. and Baldwin, T. O. Yellow light emission of Vibrio fischeri strain Y-1: Purification and characterization of the energy-accepting yellow fluorescent protein. Proc. Natl. Acad. Sci. USA 84, 8912-8916. (1987)
Desmond, E. P., Janda, J. M., Adams, F. I., and Bottone, E. J. Comparative studies and laboratory diagnosis of Vibrio vulnificus, an invasive Vibrio sp. J. Clin. Microbiol. 19, 122-125. (1984)
Devi, S. J., Hayat, U., Frasch, C. E. Kreger, A. S. and Morris, J. G. Jr. Capsular polysaccharide-protein conjugate vaccines of carbotype 1 Vibrio vulnificus: construction, immunogenicity, and protective efficacy in a murine model. Infect. Immun. 63, 2906-2911. (1995)
Devi, S. J., Hayat, U., Powell, J. L., and Morris, Jr. J. G. Preclinical immunoprophylactic and immunotherapeutic efficacy of antisera to capsular polysaccharide-tetanus toxoid conjugate vaccines of Vibrio vulnificus. Infect. Immun. 64, 2220-2224. (1996)
Engebrecht, J., Nealson, K. H. and Silverman, M. Bacterial bioluminescence: isolation and genetic analysis of functions from Vibrio fischeri. Cell 32, 773-781. (1983)
Fan, J. J., Shao, C. P., Ho, Y. C., Yu, C. K. and Hor. L. I. Isolation and characterization of a Vibrio vulnificus mutant deficient in both extracellular metalloprotease and cytolysin. Infect. Immun. 69, 5943-5948. (2001)
Farmer, J. J., Jorgensen III, J. H., Grimont, P. A. D., Akhurst, R. J., Poinar, G. O., Ageron, Jr., E., Pierce, G. V., Smith, J. A., Carter, G. P., Wilson, K. L. and Hickman-Brenner, F. W. Xenorbabdus luminescens (DNA hybridization group 5) from human clinical specimens. J. Clin. Microbiol. 27, 1594-1600. (1989)
Fleischmann, R. D., Adams, M. D., White, O., Clayton, R. A., Kirkness, E. F., Kerlavage, A. R., Bult, C. J., Tomb, J. F., Dougherty, B. A., Merrick, J. M., Mckenney, K., Sutton, G., Fitzhugh, W., Fields, C. A., Gocayne, J. D., Shirley, R., Liu, L. I., Glodek, A., Kelley, J. M., Weidman, J. F., Phillips, C. A., Spriggs, T., Hedblom, E., Cotton, M. D., Utterback, T. R., Hanna, M. C., Nguyen, D. T., Saudek, D. M., Brandon, R. C., Fine, L. D., Fritchman, J. L., Fuhrmann, J. L., Geoghagen, N. S. M., Gnehm, C. L., Mcdonald, L. A., Small, K. V., Fraser, C. M., Smith, H. O. and Venter, J. C. Whole-genome random sequencing and assembly of Haemophilus influenzae Rd. Science 269, 496-512. (1995)
Food and Nutrition Board - Institute of Medicine. Ahmed (ed), F. E. Seafood Safety/Committee on Evaluation of the Safety of Fishery Products. National Academy Press, Washington, D.C. (1991)
Foran, D. R. and Bromn, W. M. Nucleotide sequence of the luxA and luxB genes of the bioluminescent marine bacterium Vibrio fischeri. Nucleic Acids Res. 16, 777. (1988)
Frenken, L. G. J., Bos, J. W., Visser, C., Müller, W., Tommassen, J. and Verrips, C. T. An accessory gene, lipB, required for the production of active Pseudomonas glumae lipase. Mol. Microbiol. 9, 579-589. (1993)
Gaillard, J. L., Berche, P., Frehel, C., Gouin, E. and Cossart, P. Entry of L. monocytogenes into cells is mediated by internalin, a repeat protein reminiscent of surface antigens from gram-positive cocci. Cell 65, 1127-1141. (1991)
Gander, R. M. and LaRocco, M. T. Detection of piluslike structures on clinical and environmental isolates of Vibrio vulnificus. J. Clin. Microbiol. 27, 1015-1021. (1989)
Ghosh, D., Weeks, C. M., Grochulski, P., Duax, W. L., Erman, M., Rimsay, R. L. and Orr, J. C. Three-dimensional structure of holo 3α,20β-hydroxysteroid dehydrogenase: A member of a short-chain dehydrogenase family. Proc. Natl. Acad. Sci. U.S.A. 88, 10064-10068. (1991)
Gray, L. D. and Kreger, A. S. Mouse skin damage caused by cytolysin from Vibrio vulnificus and by V. vulnificus infection. J. Infect. Dis. 155, 236-241. (1987)
Gulig, P. A., Bourdage, K. L. and Starks, A. M. Molecular pathogenesis of Vibrio vulnificus. J. Microbiol. 43, 118-131. (2005)
Hada, H. S., Stemmler, J., Grossbard, M. L., West, P. A., Potrikus, C. J., Hasting, J. W. and Colwell, R. R. Characterization of non-serovar Vibrio cholerae ( Vibrio albensis) Syst. Appl. Microbiol. 6, 203-209. (1985)
Halow, K. D., Harner, R. C. and Fontenelle, L. J. Primary skin infections secondary to Vibrio vulnificus: the role of operative intervention. J. Am. Coll. Surg. 183, 329-334. (1996)
Hastings, J. W. Chemistries and colors of bioluminescent reactions: a review. Gene 173, 5-11. (1996)
Heim, R., Prasher, D. C. and Tsien, R. Y. Wavelength mutations and posttranslational autoxidation of green fluorescent protein. Proc. Natl. Acad. Sci. USA 91, 12501-12504. (1994)
Helms, S. D., Oliver, J. D., and Travis, J. C. Role of heme compounds and haptoglobin in Vibrio vulnificus pathogenicity. Infect. Immun. 45, 345-349. (1984)
Higgins, D. G. and Sharp, P. M. CLUSTAL: a package for performing multiple sequence alignment on a microcomputer. Gene 73, 237-244. (1988)
Hiroyuki, S., Hiroaki, K. and Jun’ichi O. Molecular properties and activity of amino-terminal truncated forms of lipase activator protein. Biosci. Biotechnol. Biochem. 62, 354-357. (1998)
Hlady, W. G. and Klontz, K. C. The epidemiology of Vibrio infections in Florida, 1981-1993. J. Infect. Dis. 173, 1176-1183. (1996)
Hlady, W. G., Mullen, R. C. and Hopkin, R. S. Vibrio vulnificus from raw oysters. Leading cause of reported deaths from foodborne illness in Florida. J. Fla. Med. Assoc. 80, 536-538. (1993)
Hobson, A. H., Buckley, C. M., Aamand, J. L., Jorgensen, S. T., Diderichsen, B. and McConnell, D. J. Activation of a bacterial lipase by its chaperone, Proc. Natl. Acad. Sci. U. S. A. 90(12), 5682-5686. (1993)
Ho, S. N., Horton, R. M., Pullen, J. K. and Pease, L. R. Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 77, 61-68. (1989)
Hsueh, P. R., Lin, C. Y., Tang, H. J., Lee, H. C., Liu, J. W., Liu, Y. C. and Chuang, Y. C. Vibrio vulnificus in Taiwan. Emerg. Infect. Dis. 10, 1363-1368. (2004)
Ihara, F., Kageyama, Y., Hirata, M., Nihira, T. and Yamada, Y. Purification, characterization, and molecular cloning of lactonizing lipase from Pseudomonas species. J. Biol. Chem. 266(27), 18135-18140. (1991)
Inouye, S. and Tsuji, F. I. Aequorea green fluorescent protein: Expression of the gene and fluorescence characteristics of the recombinant protein. FEBS Lett. 341, 277-280. (1994)
Jaeger, K. E., Dijkstra, B. W. and Reetz, M. T. Bacterial biocatalysts: Molecular Biology, three-dimensional structures, and biotechnological applications of lipases. Annu. Rev. Microbiol. 53, 315-351. (1999)
Jeong, K. C., Jeong, H. S., Rhee, J. H., Lee, S. E., Chung, S. S., Starks, A. M., Escudero, G. M., Gulig, P. A. and Choi, S. H. Construction and phenotypic evaluation of a Vibrio vulnificus vvpE mutant for elastolytic protease. Infect. Immun. 68(9), 5096-5106. (2000)
Johnson, D. E. and Calia, F. M. Hemolytic reaction of clinical and environmental strains of Vibrio vulnificus. J. Clin. Microbiol. 14, 457-459. (1981)
Johnston, T. C., Thompson, R. B. and Baldwin, T. O. Nucleotide sequence of the luxB gene of Vibrio harveyi and the complete amino acid sequence of the β subunit of bacterial luciferase. J. Biol. Chem. 261, 4805-4811. (1986)
Jörnvall, H., Persson, B., Krook, M., Atrian, S., Gonzalez-Duarte, R., Jeffery, J. and Ghosh, D. Short-chain dehydrogenases / reductases (SDR). Biochem. 34, 6003-6013. (1995)
Kang, M. K., Jhee, E. C., Koo, B. S., Yang, J. Y., Park, B. H., Kim, J. S., Rho, H. W., Kim, H. R. and Park, J. W. Induction of nitric oxide synthase expression by Vibrio vulnificus cytolysin. Biochem. Biophys. Res. Commun. 290, 1090-1095. (2002)
Karatani, H., Wilson, T. and Hastings, W. A blue fluorescent protein from a yellow-emitting luminous bacterium. Photochem. and Photobiol. 55, 293-299. (1992)
Kim, Y., Kim, B., Jeon, J., Kim, S., Lee, S. and Rhee, J. H. Analysis of the potential functional domains and identification of host binding partners of Vibrio vulnificus RtxA toxin. Abstr. B-142, Abstr. Annu. Meet. Am. Soc. Microbiol. (2004)
Kim, Y. R. and Rhee, J. H. Flagellar basal body flg operon as a virulence determinant of Vibrio vulnificus. Biochem. Biophys. Res. Commun. 304, 405-410. (2003)
Kok, R. G., van Thor, J. J., Nugteren-Roodzant, I. M., Brouwer, M. B. W., Egmond, M. R., Nudel, C. B., Vosman, B. and Hellingwerf, K. J. Characterization of the extracellular lipase, LipA, of Acinetobacter calcoaceticus BD413 and sequence analysis of the cloned structural gene. Molecular Microbiol. 15, 803-818. (1995)
Kok, R. G., van Thor, J. J., Nugteren-Roodzant, I. M., Vosman, B. and Hellingwerf, K. J. Characterization of lipase-deficient mutants of Acinetobacter calcoaceticus BD413: identification of a periplasmic lipase chaperone essential for the production of extracellular lipase. J. Bacteriol. 177 (11), 3295-3307. (1995)
König, B., Jaeger, K.-E., Sage, A. E., Vasil, M. L. and König, W. Role of Pseudomons aeruginosa Lipase in inflammatory mediator release from human inflammatory effector cells (platelets, granulocytes, and monocytes). Infect. Immun. 64(8), 3252-3258. (1996)
Kothary, M. H. and Kreger, A. S. Purification and characterization of an elastolytic protease of Vibrio vulnificus. J. Gen. Microbiol. 133, 1783-1791. (1987)
Kreger, A. and Lockwood, D. Detection of extracellular toxin(s) produced by Vibrio vulnificus. Infect. Immun. 33, 583- 590. (1981)
Kumamoto, K. S. and Vukich, D. J. Clinical infections of Vibrio vulnificus: a case report and review of the literature. J. Emerg. Med. 16, 61-66. (1998)
Kwon, K. B., Yang, J. Y., Ryu, D. G., Rho, H. W., Kim, J. S., Park, J. W., Kim, H. R. and Park, B. H. Vibrio vulnificus cytolysin induces superoxide anion-initiated apoptotic signaling pathway in human ECV304 cells. J. Biol. Chem. 276, 47518-47523. (2001)
Laemmli, U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685. (1970)
Lally, E. T., Hill, R. B., Kieba, I. R. and Korostoff, J. The interaction between RTX toxins and target cells. Trends Microbiol. 7, 356-361. (1999)
Lee, J. H., Rho, J. B., Park, K. J., Kim, C. B., Han, Y. S., Choi, S. H., Lee, K. H. and Park, S. J. Role of flagellum and motility in pathogenesis of Vibrio vulnificus. Infect. Immun. 72, 4905- 4910. (2004a)
Lee, S. E., Ryu, P. Y., Kim, S. Y., Kim, Y. R., Koh, J. T., Kim, O. J., Chung, S. S., Choy, H. E. and Rhee, J. H. Production of Vibrio vulnificus hemolysin in vivo and its pathogenic significance. Biochem. Biophys. Res. Commun. 324, 86-91. (2004b)
Lee, Y. R., Park, K. H., Lin, Z. Z., Kho, Y. J., Park, J. W., Rho, H. W., Koo, B. S., Kim, H. R., Song, E. K., Yu, H. N., Han, M. K., Lee, S. O., Jhee, E. C. and Kim, J. S. A calcium-calmodulin antagonist blocks experimental Vibrio vulnificus cytolysin-induced lethality in an experimental mouse model. Infect. Immun. 72, 6157- 6159. (2004c)
Linkous, D. A. and Oliver, J. D. Pathogenesis of Vibrio vulnificus. FEMS Microbiol. Lett. 174(2), 207-214. (1999)
Litwin, C. M., Rayback, T. W. and Skinner, J. Role of catechol siderophore synthesis in Vibrio vulnificus virulence. Infect. Immun. 64(7), 2834-2838. (1996)
McClure, W. R. Mechanism and control of transcription initiation in Prokaryotes. Annu. Rev. Biochem. 54, 171-204. (1985)
Meighen, E. A. Enzymes and genes from the lux operons of bioluminescent bacteria. Ann. Rev. Microbiol. 42, 151-176. (1988)
Mereno, M. L. and Landgra, M. Virulence factors and pathogenicity of Vibrio vulnificus strains isolated from seafood. Appl. Microbiol. 84(5), 747-751. (1998)
Milton, D. L., Hardman, A., Camara, M., Chhabra, S. R., Bycroft, B. W., Stewart, G. S. A. B. and Williams, P. Quorum sensing in Vibrio anguillarum: characterization of the vanI/vanR locus and identification of the autoinducer N-(3-oxodecanoyl)-L-homoserine lactone. J. Bacteriol. 179, 3004-3012. (1997)
Miyoshi, S. and Shinoda, S. Role of the protease in the permeability enhancement by Vibrio vulnificus. Microbiol. Immunol. 32, 1025-1032. (1988)
Miyoshi, S. and Shinoda, S. Activation mechanism of human Hageman factor-plasma kallikrein- kinin system by Vibrio vulnificus metalloprotease. FEBS Lett. 308, 315-319. (1992)
Molla, A., Yamamoto, T., Akaike, T., Miyoshi, S. and Maeda, H. Activation of hageman factor and prekallikrein and generation of kinin by various microbial proteinases. J. Biol. Chem. 264, 10589-10594. (1989)
Morris, J. G., Jr. and Black, R. E. Cholera and other vibrioses in the United States. N. Engl. J. Med. 312, 343-350. (1985)
Musher, D. M. Cutaneous manifestations of bacterial sepsis. Hosp. Pract. (Off. Ed). 24, 71-5, 80-2, 92 passim. (1989)
Nakajima, K., Hashimoto, T. and Yamada, Y. Two tropinone reductases with different stereospecificities are short-chain dehydrogenases evolved from a common ancestor. Proc. Natl. Acad. Sci. USA 90, 9591-9595. (1993)
Nakanishi, M., Deyashiki, Y., Nakayama, T., Sato, K. and Hara, A. Cloning and sequence analysis of a cDNA encoding tetrameric carbonyl reductase of pig lung. Biochem. Biophys. Res. Commun. 194, 1311-1316. (1993)
Nakanishi, M., Deyashiki, Y., Ohshima, K. and Hara, A. Cloning, expression and tissue distribution of mouse tetrameric carbonyl reductase. Identity with an adipocyte 27-kDa protein. Eur. J. Biochem. 228, 381-387. (1995)
Nakanishi, M., Kakumoto, M., Matsuura, K., Deyaahiki, Y., Tanaka, N., Nonaka, T., Mitsui, Y. and Hara, A. Involvement of two basic residues (Lys-17 and Arg-39) of mouse lung carbonyl reductase in NADP(H)-binding and fatty acid activation: Site-directed mutagenesis and kinetic analyses. J. Biochem. 120, 257-263. (1996)
Nardini, M., Lang, D. A., Liebeton, K., Jaeger, K.-E. and Dijkstra, B. W. Crystal structure of Pseudomons aeruginosa lipase in the open conformation. J. Biol. Chem. 275(40), 31219-31225. (2000)
Nealson, K. H. and Hastings, J. W. Bacterial bioluminescence: Its control and ecological significance. Microbiol. Rev. 43, 496-518. (1979)
Nishina, Y., Miyoshi, S., Nagase, A. and Shinoda, S. Significant role of an exocellular protease in utilization of heme by Vibrio vulnificus. Infect. Immun. 60, 2128-2132. (1992)
Noriko, O. H., Kazuhiro, Y., Takaaki, N. and Jun’ichi O. Lipase from Pseudomons aeruginosa production in Escherichia coli and activation in vitro with a protein from the downstream gene. Eur. J. Biochem. 215, 239-246. (1993)
Obeid, J. and White, P. C. Tyr-179 and Lys-183 are essential for enzymatic activity of 11β-hydroxysteroid dehydrogenase. Biochem. and Biophys. Res. Commun. 188, 222-227. (1992)
Ogierman, M. A., Fallarino, A., Riess, T., Williams, S. G., Attridge, S. R. and Manning, P. A. Characterization of the Vibrio cholera E1 Tor lipase operon lipAB and a protease gene downstream of the hly region. J. Bacteriol. 179(22), 7072-7080. (1997)
O’Kane, D. J., Karle, V. A. and Lee, J. Purification of lumazine protein from Photobacterium leiognathi and Photobacterium phosphoreum: Bioluminescence properties. Biochem. 24, 1461-1467. (1985)
Okujo, N., Akiyama, T., Miyoshi, S., Shinoda, S. and Yamamoto, S. Involvement of vulnibactin and exocellular protease in utilization of transferrin- and lactoferrin-bound iron by Vibrio vulnificus. Microbiol. Immunol. 40, 595-598. (1996)
Oliver, J. D., Roberts, D. M., White, V. K., Dry, M. A. and Simpson, L. M. Bioluminescence in a strain of the human pathogenic bacterium Vibrio vulnificus. Appl. Environ. Microbiol. 52, 1209-1211.(1986)
Oliver, J. D., Wear, J. E., Thomas, M. B., Warner, M. and Linder, K. Production of extracellular enzymes and cytotoxicity by Vibrio vulnificus. Diagn. Microbiol. Infect. Dis. 5, 99-111. (1986)
Oppermann, U.C.T., Netter, K. J. and Maser, E. Cloning and primary structure of murine 11βhydroxysteroid dehydrogenase/microsomal carbonyl reductase. Eur. J. Biochem. 277, 202-208. (1995)
Paranjpye, R. N., Lara, J. C., Pepe, C. M. and Strom, M. S. The type IV leader peptidase/N-methyltransferase of Vibrio vulnificus controls factors required for adherance to HEp-2 cells and virulence in iron-overloaded mice. Infect. Immun. 66(12), 5659-5668. (1998)
Perez-Redondo, R., Rodriguez-Garcia, A., Martin, J. F. and Liras, P. The claR gene of Streptomyces clavuligerus, encoding a LysR-type reguratory protein controlling clavulanic acid biosynthesis, is linked to the clavulanate-9-aldehyde reductase (car) gene. Gene 211, 311-321. (1998)
Prasher, D. C., Eckenrode, V. K., Ward, W. W., Prendergast, F. G. and Cormier, M. J. Primary structure of Aequorea victoria green-fluorescent protein. Gene. 111, 229-233. (1992)
Reetz M. T. and Jaeger, K-E. Overexpression, immobilization and biotechnological application of Pseudomons lipases. Chem. Phys. Lipids 93, 3-14. (1998)
Rhee, J., Kim, Y., Kim, S., Kim, C., Ryu, P., Choy, H. and Chung, S. Vibrio vulnificus Rtx toxin, a newly discovered key virulence factor, causes cytoskeletal rearrangement of host cells. Abstr. B-285, p. 81. Abstr. Annu. Meet. Am. Soc. Microbiol. (2002)
Rollof, J., Braconier, J. H., Soderstrom, C. and Nilsson-ehle, P. Interference of Staphylococcus aureus lipase with human granulocyte function. Eur. J. Clin. Microbiol. Infect. Dis. 7, 505-510. (1988)
Ross, W., Gosink, K. K., Salomon, J., Igarashi, K., Zou, C. Ishihama, A., Severionv, K. and Gourse, R. L. A third recognition element in bacterial promoters: DNA binding by the αsubunit of RNA polymerase. Science 262. 1407-1413. (1993)
Sambrook, J., Fritsch, E. F. and Maniatis, T. Molecular cloning: a laboratory manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N. Y. (1989)
Shao C. P. and Hor, L. I. Metalloprotease is not essential for Vibrio vulnificus virulence in mice. Infect. Immun. 68(6), 3569-3573. (2000)
Shryock, T. R. and Kapral, F. A. The production of bactericidal fatty acid from glycerides in staphylococcal abscesses. J. Med Microbiol. Apr, 36(4), 288-292. (1992)
Simpson, L. M. and Oliver, J. D. Siderophore production by Vibrio vulnificus. Infect. Immun. 41, 644-649. (1983)
Simpson, L. M., White, V. K., Zane, S. F. and Oliver, J. D. Correlation between virulence and colony morphology in Vibrio vulnificus. Infect. Immun. 55, 269-272. (1987)
Small, E. D., Koka, P. and Lee, J. Lumazine protein from the bioluminescent bacterium Photobacterium phosphoreum. J. Biol. Chem. 25, 8804-8810. (1980)
Smith, G. C. and Merkel, J. R. Collagenolytic activity of Vibrio vulnificus: potential contribution to its invasiveness. Infect. Immun. 35, 1155-1156. (1982)
Stormo, G. D., Schneider, T. D. and Gold, L. M. Characterization of translational initiation sites in E. coli. Nucleic Acids Res. 10, 2971-2996. (1982)
Strom, M. S. and Paranjpye, R. N. Epidemiology and pathogenesis of Vibrio vulnificus. Microbes Infect. 2, 177-188. (2000)
Studier, F. W. and Moffat, B. A. Use of bacteriophage T4 RNA polymerase to direct selective high-level expression of cloned genes. J. Mol. Biol. 189, 113-130. (1986)
Sullivan, E. R., Leahy, J. G. and Colwell, R. R. Cloning and sequence analysis of the lipase and lipase chaperone-encoding genes from Acinetobacter calcoaceticus RAG-1, and redefinition of a proteobacterial lipase family and an analogous lipase chaperone family. Gene 230 (2), 277-285. (1999)
Tison, D. L. and Kelly, M. T. Vibrio species of medical importance. Diagn. Microbiol. Infect. Dis. 2, 263-276. (1984)
Tommassen, J., Filloux, A., Bally, M., Murgier, M. and Lazdunski, A. Protein secretion in Pseudomons aeruginosa. FEMS Microbiol. Rev. 103, 73-90. (1992)
Vidal-Cros, A., Viviani, F., Boccara, M. and Gaudry, M. Polyhydroxynaphthalene reductase involved in melanin biosynthesis in Magnaporthe grisca. Eur. J. Biochem. 219, 985-992.(1994)
Vieira, J. and Messing, J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19, 259-268. (1982)
Vollberg, C. M. and Herrera, J. L. Vibrio vulnificus infection: an important cause of septicemia in patients with cirrhosis. South. Med. J. 90, 1040-1042. (1997)
Weinberg, E. D. Microbial pathogens with impaired ability to acquire host iron. Biometals. 13, 85-89. (2000)
Wermuth, B., Mader-Heinmenn, G. and Ernst, E. Cloning and expression of carbonyl reductase from rat testis. Eur. J. Biochem. 228, 473-479. (1995)
Wohlfarth, S., Hoesche, C., Strunk, C. and Winkler, U. K. Molecular genetics of the extracellular lipase of Pseudomonas aeruginosa PAO1 J. Gen. Microbiol. 138 (Pt 7), 1325-1335. (1992)
Wright, A. C. and Morris, Jr. J. G. The extracellular cytolysin of Vibrio vulnificus: inactivation and relationship to virulence in mice. Infect. Immun. 59, 192-197. (1991)
Wright, A. C., Powell, J. L., Tanner, M. K., Ensor, L. A., Karpas, A. B., Morris Jr., J. G. and Sztein, M. B. Differential expression of Vibrio vulnificus capsular polysaccharide. Infect. Immun. 67, 2250-2257. (1999)
Wright, A. C., Simpson, L. M., and Oliver, J. D. Role of iron in the pathogenesis of Vibrio vulnificus infections. Infect. Immun. 34, 503-507. (1981)
Wright, A. C., Simpson, L. M., Oliver, J. D. and Morris, Jr. J. G. Phenotypic evaluation of acapsular transposon mutants of Vibrio vulnificus. Infect. Immun. 58, 1769-1773. (1990)
Yanisch-perron, C., Vieira, J. and Messing, J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33, 103-119. (1985)
Yoshida, S., Ogawa, M. and Mizuguchi, Y. Relation of capsular materials and colony opacity to virulence of Vibrio vulnificus. Infect. Immun. 47, 446-451. (1985)
Young, G. M., Schmiel, D. H., and Miller, V. L. A new pathway for the secretion of virulence factors by bacteria: the flagellar export apparatus functions as a protein-secretion system. Proc. Natl. Acad. Sci. U. S. A. 96, 6456-6461. (1999)
Zuppardo, A. B. and Siebeling, R. J. An epimerase gene essential for capsule synthesis in Vibrio vulnificus. Infect. Immun. 66(6), 2601-2606. (1998)