Gene cloning and prokaryotic expression of recombinant flagellin A from Vibrio parahaemolyticus

Ye Yuan; Xiuli Wang; Sheping Guo; Yang Liu; Hui Ge; Xuemei Qiu

doi:10.1007/s00343-010-0068-1

Your Location：

Home >

Browse articles >

Gene cloning and prokaryotic expression of recombinant flagellin A from Vibrio parahaemolyticus

Updated：2023-05-16

- Gene cloning and prokaryotic expression of recombinant flagellin A from Vibrio parahaemolyticus
- Journal of Oceanology and Limnology Vol. 28, Issue 6, Pages: 1254-1260(2010)
- Affiliations：
  
  College of Life Sciences and Biotechnology Dalian Fisheries University
- Author bio：
- Funds：
- DOI：10.1007/s00343-010-0068-1
  CLC：
- Online First：24 June 2010，
  
  Published：2010
- Accepted：
Scan QR Code
Yuan, Y., Wang, X., Guo, S. et al. Gene cloning and prokaryotic expression of recombinant flagellin A from Vibrio parahaemolyticus, ., Chin. J. Ocean. Limnol. 28, 1254–1260 (2010). https://doi.org/10.1007/s00343-010-0068-1
DOI：

Yuan, Y., Wang, X., Guo, S. et al. Gene cloning and prokaryotic expression of recombinant flagellin A from Vibrio parahaemolyticus, ., Chin. J. Ocean. Limnol. 28, 1254–1260 (2010). https://doi.org/10.1007/s00343-010-0068-1 DOI：

摘要

Abstract

The Gram-negative Vibrio parahaemolyticus is a common pathogen in humans and marine animals. Bacteria flagellins play an important role during infection and induction of the host immune response. Thus

flagellin proteins are an ideal target for vaccines. We amplified the complete flagellin subunit gene (flaA) from V. parahaemolyticus ATCC 17802. We then cloned and expressed the gene into Escherichia coli BL21 (DE3) cells. The gene coded for a protein that was 62.78 kDa. We purified and characterized the protein using Ni-NTA affinity chromatography and Anti-His antibody Western blotting

respectively. Our results provide a basis for further studies into the utility of the FlaA protein as a vaccine candidate against infection by Vibrio parahaemolyticus. In addition

the purified FlaA protein can be used for further functional and structural studies.

关键词

Keywords

references

Arora S K, Ritchings B W, Almira E C, Lory S, Ramphal R. 1997. A transcriptional activator, FleQ, regulates mucin adhesion and flagellar gene expression in Pseudomonas aeruginosa in a cascade manner. J. Bacteriol., 179(17): 5 574–5 581.

Brandfass C, Karlovsky P. 2008. Upscaled CTAB-based DNA extraction and real-time PCR assays for Fusarium culmorum and F. graminearum DNA in plant material with reduced sampling error. Int. J. Mol. Sci., 9(11): 2 306–2 321.

Chávez A, García-Huante Y, Ruiz B, Langley E, Rodríguez-Sanoja R. Sanchez S. 2009. Cloning and expression of the sco2127 gene from Streptomyces coelicolor M145. J. Ind. Microbiol. Biotechnol., 36(5): 649–654.

Das M, Chopra A K, Wood T, Peterson J W. 1998. Cloning, sequencing and expression of the flagellin core protein and other genes encoding structural proteins of the Vibrio cholerae flagellum. FEMS Microbiol. Lett., 165(2): 239–246.

Datta S, Janes M E, Simonson J G. 2008. Immunomagnetic separation and coagglutination of Vibrio parahaemolyticus with anti-flagellar protein monoclonal antibody. Clin. Vaccine. Immunol., 15(10): 1 541–1 546.

Fan J F, Song L C, Wang B, Zang H M, Liang Y B. 2006. Preliminary study on one pathogenic bacterium-Vibrio parahaemolyticus associated with red body disease in Litopenaeus vannamei. Marine Sciences, 30(4): 40–44. (in Chinese with English abstract)

Jacchieri S G, Torquato R, Brentani R R. 2003. Structural study of binding of flagellin by Toll-like receptor 5. J. Bacteriol., 185(14): 4 243–4 247.

Kimura M. 1980. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J. Mol. Evol., 16(2): 111–120.

Klose K E, Mekalanos J J. 1998. Differential regulation of multiple flagellins in Vibrio cholerae. J. Bacteriol., 180(2): 303–316.

Lee J H, Rho J B, Park K J, Kim C B, Han Y S, Choi S H, Lee K H, Park S J. 2004. Role of flagellum and motility in pathogenesis of Vibrio vulnificus. Infect. Immun., 72(8): 4 905–4 910.

Lee K K, Liu P C, Huang C Y. 2003. Vibrio parahaemolyticus infectious for both humans and edible mollusk abalone. Microbes. Infect, 5(6): 481–485.

Liu Z, Miyashiro T, Tsou A, Hsiao A, Goulian M, Zhu J. 2008. Mucosal penetration primes Vibrio cholerae for host colonization by repressing quorum sensing. Proc. Natl. Acad. Sci. USA., 105(28): 9 769–9 774.

Macnab R M. 2003. How bacteria assemble flagella. Annu. Rev. Microbiol., 57: 77–100.

McCarter L L. 2001. Polar flagellar motility of the Vibrionaceae. Microbiol. Mol. Biol. Rev., 65(3): 445–462.

McGee K, Hörstedt P, Milton D L. 1996. Identification and characterization of additional flagellin genes from Vibrio anguillarum. J Bacteriol., 178(17): 5 188–5 198.

Millikan D S, Ruby E G. 2004. Vibrio fischeri flagellin A is essential for normal motility and for symbiotic competence during initial squid light organ colonization. J. Bacteriol., 186(13): 4 315–4 325.

Milton D L, O’Toole R, Hörstedt P, Wolf-Watz H. 1996. Flagellin A is essential for the virulence of Vibrio anguillarum. J. Bacteriol., 178(5): 1 310–1 319.

Moorthy S, Watnick P I. 2004. Vibrio fischeri flagellin A is essential for normal motility and for symbiotic competence during initial squid light organ colonization. J. Bacteriol., 186(13): 4 315–4 325.

Ormonde P, Hörstedt P, O’Toole R, Milton D L. 2000. Role of motility in adherence to and invasion of a fish cell line by Vibrio anguillarum. J. Bacteriol., 182(2): 2 326–2 328.

Qian R, Xiao Z, Zhang C, Chu W, Mao Z, Yu L. 2008. Expression and purification of two major outer membrane proteins from Vibrio alginolyticus. World. J. Microbiol. Biotechnol., 24: 245–251.

Qin Y X, Wang J, Wang S F, Su Y Q, Zhuang X. 2007. Study on the expression of recombined plasmid pcFlaA in Epinephelus awoara. High Tech. Lett., 17(7): 761–764. (in Chinese with English abstract)

Ramos H C, Rumbo M, Sirard J C. 2004. Bacterial flagellins: mediators of pathogenicity and host immune responses in mucosa. Trends Microbiol., 12(11): 509–517.

Saitou N, Nei M. 1987. The neighbour-joining method: a new method for reconstructing phylogenetic trees. Mol. Biol. Evol., 4(4): 406–425.

Shapiro L. 1995. The bacterial flagellum: from genetic net-work to complex architecture. Cell, 80(4): 525–527.

Thompson F L, Iida T, Swings J. 2004. Biodiversity of vibrios. Microbiol. Mol. Biol. Rev., 68(3): 403–431.

Thompson J D, Higgins D G, Gibson T J. 1994. CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position specific gap penalties and weight matrix choice. Nucleic Acids Res., 22(22): 4 673–4 680.

Wu Z L, Nie J, Bao H N. 2001. Expression of thermostable direct hemolysin gene of Vibrio parahaemolyticus in E. coli and activity of expressed product. J. Cell. Mol. Immunol., 17(5): 426–427.

Young G M, Smith M J, Minnich S A, Miller V L. 1999. The Yersinia enterocolitica motility master regulatory operon, flhDC, is required for flagellin production, swimming motility, and swarming motility. J. Bacteriol., 181(9): 2 823–2 833.

Zhuang X, Qin Y X, Su Y Q, Wang J, Ding S X. 2007. Cloning and sequencing of flaA gene of Vibrio harveyi and construction of its eukaryotic expression recombinant plasmid. Acta Oceanol. Sin., 29(6): 74–79. (in Chinese with English abstract)

Zorrilla I, Arijo S, Chabrillon M, Diaz P, Martinez-Manzanares E, Balebona M C, Moriñigo M A. 2003. Vibrio species isolated from diseased farmed sole, Solea senegalensis (Kaup), and evaluation of the potential virulence role of their extracellular products. J. Fish. Dis., 26(2): 103–108.

Views

Downloads

CSCD

Alert me when the article has been cited

Submit

Tools

Publicity Resources

Gene cloning and prokaryotic expression of recombinant outer membrane protein from Vibrio parahaemolyticus

Characterization and stability of sedimentary colloids in different ecology regions in Taihu Lake

Screening and characterization of proteases produced by deep-sea cold seep bacteria

Preparation, characterization, and antifungal evaluation of a new type of aminourea chitooligosaccharide derivatives

Expression, purification, and subcellular localization of phospholipase C in Dunaliella salina

Related Author

Ye Yuan

Xiuli Wang

Sheping Guo

Xuemei Qiu

Shiyin LI

Kaipian SHI

Li YIN

Juan WANG

Related Institution

College of Life Sciences and Biotechnology Dalian Ocean University

School of Environment, Nanjing Normal University

Jiangsu Open Laboratory of Large-scale Scientific Instruments, Nanjing Normal University

Department of Geological Sciences, University of Alabama, Tuscaloosa AL

Jiangsu Center for Collaborative Innovation in Geographical Information Resource Development and Application

Address：Editorial Office of JOL, 88 Haijun Rd. Qingdao, 266000, China
Tel：+86-532-82898754 Email：jol@qdio.ac.cn
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备09064830号-19 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰