DOI QR코드

DOI QR Code

Immuno Gold 표지법을 이용한 대장균내 Vibrio fluvialis MotX 단백질의 존재 부위 결정

Detection of the Recombinant MotX Protein Vibrio fluvialis in Escherichia coli with Immuno-Gold Labeling Method

  • LEE Jong Hee (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Park Jae Hyun (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Kim Sun Hoi (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • An Sun Hee (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Kong In Soo (Department of Biotechnology and Bioengineering, Pukyong National University)
  • 발행 : 2002.07.01

초록

The rotation of the flagellar motor is powered by the electrochemical gradient of specific ions across the cytoplasmic membrane. Recently, the gents of the Na'-driven motor have been cloned from marine bacterium of Vibrio sp. and some of the motor proteins have been purified and characterized. Also, motx gene encoding a channel component of the sodium type flagellar motor was identified from Vibrio Huuiaiis (KTCC 2473). The amino acid sequence of MotX protein from V, Huvialis shared 90, 85, $85\%$ identity with V, cholerae, V. alginolyticus, V parahaemolyticus, respectively. We have studied the localization of the expressed MotX protein in Escherichia coli by immune-gold labeling of ultra-thin frozen section. Our observation of the expressed protein indicated that MotX protein could be existed as attachment to inner membrane in E. coli.

키워드

참고문헌

  1. Asai, Y., S. Kojima, H. Kato, N. Nishioka, I. Kawagishi and M. Homma. 1997. Putative channel components for the fast-rotating sodium-driven flagellar motor of a marine bacterium. J. Bacteriol., 179, 5104-5110 https://doi.org/10.1128/jb.179.16.5104-5110.1997
  2. Atsumi, T., L. McCarter and Y. Imae. 1992. Polar and lateral flagellar motors of marine Vibrio are driven by different ion-motive forces. Nature, 355, 182-184 https://doi.org/10.1038/355182a0
  3. Jaques, S., Y.K. Kim and L. McCarter. 1999. Mutations conferring resistance to phenamil and amiloride, inhibitors of sodium-driven motility of Vibrio parahaemolyticus. Proc. Natl. Acad. Sci. USA, 96, 5740-5745 https://doi.org/10.1073/pnas.96.10.5740
  4. Kawagishi, I., Y. Maekawa, T. Atsumi, M. Homma and I. Yasuo. 1995. Isolation of the polar and lateral flagellum-defective mutants in Vibiio alginolyticus and identification of their flagellar driving energy sources. J. Bacteriol., 177, 5158-5160 https://doi.org/10.1128/jb.177.17.5158-5160.1995
  5. Lee, J.V., P. Shread, L. Fumess and T.N. Bryant. 1981. Taxonomy and description of Vibrio fluvialis sp. nov. (synonymo group Fvibnos, group EF 6). J. Appl. Bactenol, 50, 73-94 https://doi.org/10.1111/j.1365-2672.1981.tb00873.x
  6. McCarter, L. 1994a. MotX, the channel component of the sodium-type flagellar motor. J. Bactehol., 176, 5988-5998
  7. McCarter, L. 1994b. MotY, a component of the sodium-type flagellar motor. J. Bacteriol., 176, 4219-4225 https://doi.org/10.1128/jb.176.14.4219-4225.1994
  8. McCarter, L. and M. Silverman. 1990. Surface-induced swanner cell differentiation of Vibrio parahaemolyticus. Mol. Microbiol., 4, 1057-1062 https://doi.org/10.1111/j.1365-2958.1990.tb00678.x
  9. Okunishi, I., I. Kawagishi and M. Homma. 1996. Cloning and characterization of motY, a gene coding for a component of the sodium-driven flagellar motor in Vibrio atginilyticus. J. Bacteriol., 178, 2409-2415 https://doi.org/10.1128/jb.178.8.2409-2415.1996
  10. Okabe, M., T. Yakushi, Y. Asai and M. Homma. 2001. Cloning and characterization of motX, a Vibrio algiiiolyticus sodium-dhven flagella motor gene. J. Biochem., 130, 879-884 https://doi.org/10.1093/oxfordjournals.jbchem.a003061
  11. Park, J.H., J.H. Lee, Y.S. Kim, Y.K. Hong and I.S. Kong. 2001. Molecular cloning and expression of a sodium-diiven flagella motor component gene (motX) from Vibrio fluvialis. J. Microbiol. Biotechnol., 11, 973-978
  12. Wagenaar, F., G.L. Kok, J.M.B. Davies and J.M.A. Pol. 1993. Rapid cold Iixation of tissue samples by microwave irradiation for use in electron microscopy. Histochem. J., 25, 719-725
  13. Ybmohiro, T. and H. Michio. 2001. $Na^+$ -driven flagella motor of Vibrio. Biochim. Biophys. Acta., 1505, 82-93 https://doi.org/10.1016/S0005-2728(00)00279-6
  14. Yorimitsu, T., K. Sato, Y. Asai, I. Kawagishi and M. Homma. 1999. Functional interaction between pomA and pomB, the sodiumdriven flagellar motor components of Vibrio alginolyticus. J. Bacteriol., 181, 5103-5106