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研究生:施鈞傑
研究生(外文):Chun-Chieh Shih
論文名稱:分別表現鼠傷寒沙門氏桿菌之yjeH及ydgFE基因導致ceftriaxone之抗藥性
論文名稱(外文):Respectively to overexpress yjeH and ydgFE genes to confer ceftriaxone resistance in Salmonella enterica serovar Typhimurium
指導教授:胡文熙
指導教授(外文):Wensi S. Hu
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:醫學生物技術研究所
學門:醫藥衛生學門
學類:醫學技術及檢驗學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:78
中文關鍵詞:鼠傷寒沙門氏桿菌抗藥性基因
外文關鍵詞:overexpressyjeHydgFEceftriaxoneresistanceSalmonella Typhimurium
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摘要
實驗室之前利用Tn5 mutagenesis的技術將對Ceftriaxone藥物具有高抗性的鼠沙門氏桿菌WT200之基因破壞,造就了transposon insertion突變株。其中編號6B7是一個transporter yjeH基因被Tn5破壞的突變株, 8F9是一個transporter ydgF基因被Tn5破壞的突變株,兩突變株皆比原來WT200對於CRO之MIC下降四倍之多。為了闡明這兩個基因是否與抵抗CRO藥物有相關性,我們構築了各別帶有yjeH基因及ydgFE基因的表現載體pBAD-yjeH-DsRed、pBAD- ydgFE-DsRed,去實行互補試驗。ydgFE是一個由ydgF及ydgE基因所組成的operon。結果顯示將pBAD-yjeH-DsRed送入6B7突變株中,將會回復與WT200對於CRO之MIC一樣,意味著在6B7突變株表現YjeH蛋白將完全回復對於CRO的抗藥性。另外,我們利用Efflux pump抑制劑CCCP來測試WT200及WT200內含pBAD-yjeH-DsRed對於CRO之MIC。結果發現,在WT200過量表現YjeH蛋白對於CRO之MIC高於沒有過量表現YjeH蛋白的WT200。這是另一個YjeH蛋白的確能賦予鼠傷寒沙門氏桿菌抵抗CRO能力的證明。因為YjeH蛋白是一個屬於APA家族的putative transporter,而且目前為止從未有人提出它與抗藥相關性的報導。在我們的研究中,是第一個証明屬於APA家族的YjeH transporter與抗藥有相關。CRO是否為這個transporter的受質之一需要在未來的研究中更進一步的證明。在本研究中同時又以互補試驗來証明ydgFE基因的確與鼠傷寒沙門氏桿菌抵抗CRO藥物有相關性。而YdgFE是一個被預測屬於Small drug resistance家族的transporter,至於ydgFE如何幫助細菌來抵抗CRO藥物將在未來的研究中更進一步的探討。
Abstract
We previously generated transposon insertion mutant which derived from a ceftriaxone(CRO)-resistant strain of Salmonella enterica serovar Typhimurium WT200. Mutant 6B7 and 8F9 are transporter genes yjeH and ydgF which truncated by transposon insertion, respectively, and both have 4-fold reduction of CRO resistance. To elucidate whether these two genes correlated with CRO resistance, we constructed the expression vectors pBAD-yjeH-DsRed and pBAD-ydgFE-DsRed which carry functional yjeH and ydgFE genes, respectively, for the complementation test. The ydgFE is composed of two genes ydgF and ydgE in the same operon. The MICs of mutant 6B7 strain with pBAD-yjeH-DsRed against CRO increased to the same level of WT200. This result indicated overexpression of YjeH can completely restore CRO resistance in 6B7 mutant strain. WT200 strain with or without pBAD-yjeH-DsRed were tested against CRO in the presence of CCCP, an energy uncoupler that has been shown to inhibit the action of efflux pumps. The MICs of WT200 strain with YjeH overexpression are higher than those of bacteria without YjeH overexpression. This is the additional evidence to suggest that YjeH transporter protein can confer CRO resistance in Salmonella typhimurium. Since YjeH is a putative transporter, which belongs to APA family, and was never reported to correlate with drug resistance. In this study, it is the first time to prove that the member of APA family is associated with drug resistance . Further studies are needed to prove whether CRO is the substrate of this transporter. We were also able to demonstrate ydgEF genes are correlated with CRO resistance in Salmonella typhimurium by the complementation test. Since YdgEF is a putative cation transporter which belongs to small drug resistance family of efflux pump system, we will perform more studies to prove how ydgEF can confer CRO resistance in the near future.
參考文獻
Alekshun, M. N., and S. B., Levy. 1997. Regulation of chromosomally mediated multiple antibiotic resistance:the mar regulon. Antimicrob. Agents. Chemoter.:41:2067-2075.
Alonso, A., G. Morales, R. Escalange, E. Campanario, L. Sastre, and J. L. Martinez. 2004 Overexpression of the multidrug efflux pump SmeDEF impairs strenostrophomonas maltophilia physiology. Journal of Antimicrobial Chemotherapy. 53:432-434.
Altekruse, S. F., M. L., Cohen, and D. L., Swerdlow. 1997. Emerging foodborne diseases. Emerg. Infect. Dis., 3:285-293.
Arthur, M., and C. Patrice. 1993 Genetics and mechanisms of glycopetide resistance in Enterococci. Antimicrobial Agents and Chemotherapy. 37(8): 1563-1571.
Bachmann, B. J. 1990. Linkage map of Escherichia coli K-12, edition 8. Microbiol. Rev 54, 130-197.
Barua, S., T. Yamashino, T. Hasegawa, K. Yokoyama, K. Torii, M. Ohta. 2002 Involvement of surface polysaccharide in the organic acid resistance of Shiga Toxin-producing Escherichia coli.O157:H7. Molecular Microbiology. 43:629-640.
Boonmar, S., A. Bangtrakulnonth, S. Pornruangwong, S. Samosornsuk, K. Kaneko, and M. Ogawa. 1998. Significant increase in antibiotic resistance of salmonella isolates from human beings and chicken meat in Thailand. Vet. Microbiol., 62:73-80.
Brown, J. M., and K. J. Shaw. 2003 A novel family of Escherichia coli Toxin-Antitoxin gene pairs. J. Bacteriol.185(22):6600-6608.
Bush, K., G. A. Jacoby, and A. A. Medeiros. 1995. A functional classification scheme for β-lactamases and its correlation with molecular structure. Antimicrobial Agents and Chemotherapy. 39:1211-1233.
Cheng, C. Y. 2003. Outer membrane protein profile in ceftriaxone resistant Salmonella typhimurium by two-dimensional gel electrophoresis analysis. MS thesis. National Yang-Ming University, Taiwan.
Chopra, I., and M. Roberts. 2001. Tetracycline antibiotics: mode of action, applications, molecular biology, and epidemiology of bacterial resistance. Microbiology and Molecular Biology Reviews, 65:232-260.
Daly, M., and S. Fanning. 2000. Characterization and Chromosomal Mapping of Antimicrobial Resistance Genes in Salmonella enterica serotype Typhimurium. Appl. Environ. Microbiol., 66:4842-4848.
Darget, M., and S. D. Ehrlich. 1979. Prolonged incubation in CaCl2 improves the competence of Escherichia coli cells. Gene, 6:23-28.
Delavega, A. L., and H. A. Delcour. 1996 Polyamines decrease Escherichia coli outer membrane permeability. J. Bacteriol.178(13):3715-3721.
Denyer, S. P. and J. Y. Maillard. 2002. Cellular impermeability and uptake of biocides and antibiotics in gram-negative bacteria.J. Applied Microbiology Symposium Supplement. 92:355-455.
Donald, L. J., M. Y. Nelson, and H. S. J. Milton. 2000 The amino acid/polyamine organocation (APC) superfamily of transporters specific for amino acid, polyamines and organocations. Microbiology 146:1797-1814.
Donald, L. J., M. Y. Nelson, and H. S. J. Milton. 2001 The drug/metabolite transporter superfamily. Eur. J. Biochem. 268:3620-3639.
Fralick, J. A. 1996. Evidence that TolC is required for functioning of the Mar/AcrAB efflux pump of Escherichia coli. J. Bacteriol. 78:5803-5805.
Fuminobu, Y., and N. Hiroshi. 1985 Diffusion of β-Lactam antibiotics through the porin channels of Escherchia coli K-12. Antimicrobial Agents and Chemotherapy.27(1):84-92.
Gilson, L., H. K. Mahantym, and R. Kolter. 1990. Genetic analysis of an MDR-like export system: the secretion of colicin V. EMBO J. 9:3875-3894.
Glynn, M. K., C. Bopp, W. Dewitt, P. Dabney, M. Mokhtar, Mokhtar, and F. J. Angulo. 1998. Emergence of multidrug resistant salmonella enterica serotype Typhimurium DT104 infections in the United States. N. Engl. J. Med., 338:1333-1338.
Grkovic, S., M. H. Brown, and R. A. Skurray. 2002 Regulation of bacterial drug export systems. Microbiology and Molecular Biology Reviews.66(4):671-707.
Gross, U., H. Tschape, I. Bednarek, and M. Frosch. 1998. Antibiotic resistance in Salmonella enterica serotype typhimurium. Eur. J. Clin. Microbiol. Infect. Dis., 17: 385-387.
Guzman, L. M., D. Belin, M. J. Carson, and J. Beckwith. 1995. Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter. J. Bacteriol. 177:4121-4130.
Hancock, R. E. W. 1984 Alterations in outer membrane permeability. Ann. Rev. Microbiol. 38:237-64.
Hancock, R. E. W. 1997. The bacterial outer membrane as a drug barrier. Trends in Microbiology.5:37-42.
Hermann, L., 2003 ABC-transporters: implications on drug resistance from microorganisms to human cancers. International Journal of antimicrobial agents. 22:188-199.
Samartzidou, H., and A. H. Delcour. 1999 Excretion of endogenous cadaverine leads to decrease in porin-mediated outer membrane permeability. J. Bacteriol.181(3):791-798.
Huang, J., W. O. Paul, S. Wei, M. Heather, J. Xinhe, L. Neethan, M. P. Leslie, V. Leroy, F. Frank, N. G. Micheal, and M. Damien. 2004 Novel chromosomally encoded multidrug efflux transporter MdeA in Staphylococcus aureus. Antimicrobial Agents and Chemotherapy. 48(3):909-917.
Johnson, J. M., and G. M. Church. 1999. Alignment and structure prediction of divergent protein families: periplasmic and outer membrane proteins of bacterial efflux pumps. J. Mol. Biol. 287: 695-715.
Kashiwagi, K., S. Shibuya, H. Tomitori, A. Kuraishi, K. Igarashi. 1997. Excretion and uptake of putrescine by the PotE protein in Escherichia coli. J Biol Chem. 272(10):6318-23.
Klein, N.C., and B.A. Cunha. 1995. The selection and use of cephalosporins: a review. Adv-Ther., 12(2):83-101.
Kobayashi, N., K. Nishino, and A. Yamaguchi. 2001.Novel macrolide- specific ABC-type efflux transporter in Escherichia coli. 183:5639-5644.
Li, P. C. 2003 Overexpression of the response regulator BaeR of the two-component system confer ceftriaxone resistance in Salmonella typhimurium. MS thesis. National Yang-Ming University, Taiwan.
Maniatis, T., E. F. Fritsch, and J. Sambrook. 1982. Molecular cloning: a laboratory manual.Cold Spring Habor Laboratory, Cold Spring Habor, New York.
Ma, D., D. N. Cook, M. Alberti, N. G, Pon, H. Nidaido, and J. E. Hearst. 1993. Molecular cloning and characterization of acrA and acrE genes of Escherichia coli. J. Bacteriol. 175:6299-6313.
Ma, D., D. N. Cook, M. Alberti, N. G. Pon, H. Nidaido, and J. E. Hearst. 1995. Genes acrA and acrB encode a stress-induced efflux system of Escherichia coli. Mol. Microbiol. 16:45-55.
McEvoy, G. K., editor. 8:12. 06 Cephalosporins. AHFS Drug Information 98. Bethesda; American Society of Health-System Pharmacists, Inc. 1998:125-205.
Mcclelland, M. et al. 2001. Complete genome sequence of Salmonella enterica serovar typhimurium LT2. Nature. 2001. 413: 852-856.
Mikhail, V. M., F. F. Arkady, A. L. Yulii, P. S. Aleksandr, G. Z. Andrey, L. M. Mikhail, and A. L. Sergey. 1999 Fluorescent proteins from nonbioluminescent Anthozoa species. Nature Biotechnology 17:969-973.
Mortimer, P. G., and L. J. Piddock. 1993. The accumulation of five antibacterial agents in porin-deficient mutants of Escherichia coli. Journal of Antimicrobial Chemotherapy, 32:195-213.
Molloy, M. P., B. R. Herbert, M. B. Slade, T. Rabilloud, A. S. Nouwens, K. L. Willians, and A. A. Gooley. 2000. Proteomic analysis of the Escherichia coli outer membrane. Eur. J. Biochem., 267, 2871-2881.
Molly, M. P., N. D. Phadke, J. R. Maddock, and P. C. Andrews. 2001. Two-dimensional electrophoresis and peptide mass fingerprinting of bacterial outer membrane proteins. Electrophoresis, 22:1686-1696.
Monique, P., W. V. Hendrik, and W. N. Konings. 2000 Molecular Properties of Bacterial Multidrug Transporters. Microbiology and Molecular Biology Reviews. 64(4):672-693.
Nikaido, H. 1994. Prerention of drug access to bacterial targets: permeability barriers and active efflux. Science. 264:382-388.
Nikaido, H. 1998. Antibiotic resistance caused by gram-negative multudrug efflux pumps. Clin. Infect. Dis. 27: S32-S41.
Nikaido, H. 1998. Multiple antibiotic resistance and efflux. Current Opinion in Microbiology. 1:516-523.
Nikaido, H., M. Basina, V. Nauyen, and E. Y. Rosenberg. 1998 Multidrug efflux pump AcrAB of Salmonella typhimurium excretes only those β-Lactam antibiotics contaning lipophilic side chains J. Bacteriol. 180(17):4686-4692.
Nikaido, H., E. Y. Rosenberg, and J. Foulds. 1983. Porin channels in Escherichia coli: studies with beta-lactams in intact cells. Journal of Bacteriology, 153:232-240.
Nikaido, H. and H. I. Zgurskaya. 1999. Antibiotic efflux mechanisms. Curr. Opin. Infect. Dis. 12 529-536.
Nikaido, H., and H. I. Zgurskaya. 2001. AcrAB and related multidrug efflux pumps of Escherichia coli. J. Mol. Microbiol. Biotechnol., 3:215-218.
Nishino, K., and A. Yamaguchi. 2001 Analysis of a complete library of putative drug transporter genes in Escherichia coli. J. Bacteriaol. 183:5803-5812.
Nikaido, H., and H. I. Zgurskaya. 2001. AcrAB and related multidrug efflux pumps of Escherichia coli. J. Mol. Microbiol. Biotechnol., 3:215-218.
Okusu, H., D. Ma , and H. Nikaido. 1996. AcrAB efflux pump plays a major role in the antiviotic resistance phenotype of Escherichia coli multiple-antibiotic-resistance phenotype (Mar) mutants. J. Bacteriaol. 178:306-308.
Pan, T. M., T. K. Wang, C. H. Lee, S. W. Chien, and C. B. Horng. 1997. Food-borne disease outbreaks due to bacteria in Taiwan, 1986 to 1995. J. Clin. Microbiol., 35:1260-1262.
Poole, K. 2002. Mechanisms of bacterial biocide and antibiotic resistance. Journal of Applied Microbiology Symposium Supplement, 92:55S-64S.
Randall, L. P., and M. J. Woodward. 2002 The multiple antibiotic resistance(mar) locus and its significance. Res Vet Sci., 72(2):87-93.
Minchin, R. F., and J. E. McCoubrie. 2004 Mutational analysis of the large periplasmic loop 7-8 of the putrescine transporter PotE in Eschericha coli. The International Journal of Biochemistry & Cell Biology. 36:271-280.
Ren, Q. et al. 2004 TransportDB: A Relational Database of Cellular Membrane Transport Systems. Nucleic Acids Research. 32: D284-D288.
Russell, A. D., and J. R. Furr. 1996. Susceptibility of porin and lipopolysaccharide-deficient strans of Escherichia coli to some antiseptics and disinfectants. Journal of Hospital Infection. 8: 47-56.
Russell, A. D. 2002 Antibiotic and biocide resistance in bacteria: Introduction. Journal of Applied Microbiology Symposium Supplement, 92:1S-3S.
Sambrook, J., E. F. Fritsch, and T. Manitis. 1989. Molecular Cloning: a laboratory manual. Cold Spring Habor Laboratory, Cold Spring Habor, New York.
Scott Snyder, D., and T. J. Mcintosh. 2000 The Lipopolysaccharide Barrier: Correlation of Antibiotic Susceptibility with Antibiotic Permeability and Fluorescent Probe Binding Kinetics. Biochemistry 39:11777-11787.
Soksawatmaekhin, W., A. Kuraishi, K. Sakata, K. Kashiwagi, K. Igarashi. 2004. Excretion and uptake of cadaverine by CadB and its physiological functions in Escherichia coli. Mol Microbiol. 51(5):1401-12.
Takiff, H. E., M. Cimino, M. C. Musso, T. Weisbrod, R. Martinez, M. B. Delgado, L. Salazar. 1996 Efflux pump of the proton antiporter family confers low-level fluoroquinolone resistance in Mycobacterium smegmatis. Proc. Natl. Acad. Sci. USA. 93: 362-366.
Thomson, K. S., and M. E. Smith. 2000. Version 2000: the new β- lactamases of gram-negative bacteria at the dawn of the new millennium.Microbes and Infection. 2:1225-1235.
Yang, Y. L., C. C. Liu, S. M. Wang, J. J. Wu, A. H. Huang, and C. P. Cheng. 1998. High rates of antimicrobial resistance among clinical isolates of nontyphoidal Salmonella in Taiwan. Eur. J. Clin. Microbiol. Infect. Dis., 17:880-883.
Yethon, J. A., J. S. Gunn, R. K. Ernst, S. I. Miller, L. Laroche, D. Malo, C. Whitfield. 2000. Salmonella enterica serovar typhimurium waaP mutants show increased susceptibility to polymyxin and loss of virulence in vivo. Infection and Immunity. 68:4485-4491.
Yoneyama, H., Y. Yamani, and T. Nakae. 1995. Role of porins in the antiviotic susceptibility of Pseudomonas aeruginosa construction of mutants with delections in the multiple porin genes. Biochemical and Biophysical Research Communications. 213:88-95.
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