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

Ye Yuan; Xiuli Wang; Sheping Guo; Xuemei Qiu

doi:10.1007/s00343-011-0211-7

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

OriginalPaper | Updated：2023-05-16

- Gene cloning and prokaryotic expression of recombinant outer membrane protein from Vibrio parahaemolyticus
- Gene cloning and prokaryotic expression of recombinant outer membrane protein from Vibrio parahaemolyticus
- 海洋湖沼学报（英文） 2011年29卷第5期页码：952-957
- Affiliations：
  
  College of Life Sciences and Biotechnology Dalian Ocean University
- Author bio：
- Funds：
  
  Supported by the Key Laboratory Foundation of the Educational Department of Liaoning Province (No. 2009S024), the Dalian Municipal Government of China (No. 2007B11NC069), and the Grant of Dalian Ocean University (No. SY2007005)
- DOI：10.1007/s00343-011-0211-7
  中图分类号：
- 网络首发：2011-06-22，
  
  纸质出版：2011
- Accepted：
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Gene cloning and prokaryotic expression of recombinant outer membrane protein from Vibrio parahaemolyticus[J]. 海洋湖沼学报（英文）, 2011,29(5):952-957.

Yuan, Y., Wang, X., Guo, S. et al. Gene cloning and prokaryotic expression of recombinant outer membrane protein from Vibrio parahaemolyticus, ., Chin. J. Ocean. Limnol. 29, 952–957 (2011). https://doi.org/10.1007/s00343-011-0211-7
Gene cloning and prokaryotic expression of recombinant outer membrane protein from Vibrio parahaemolyticus[J]. 海洋湖沼学报（英文）, 2011,29(5):952-957. DOI： 10.1007/s00343-011-0211-7.

Yuan, Y., Wang, X., Guo, S. et al. Gene cloning and prokaryotic expression of recombinant outer membrane protein from Vibrio parahaemolyticus, ., Chin. J. Ocean. Limnol. 29, 952–957 (2011). https://doi.org/10.1007/s00343-011-0211-7 DOI：

摘要

Abstract

Gram-negative Vibrio parahaemolyticus is a common pathogen in humans and marine animals. The outer membrane protein of bacteria plays an important role in the infection and pathogenicity to the host. Thus

the outer membrane proteins are an ideal target for vaccines. We amplified a complete outer membrane protein gene (ompW) 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 42.78 kDa. We purified the protein using Ni-NTA affinity chromatography and Anti-His antibody Western blotting

respectively. Our results provide a basis for future application of the OmpW protein as a vaccine candidate against infection by V. parahaemolyticus. In addition

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

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references

Albrecht R, Zeth K, Soding J, Lupas A, Linke D. 2006. Expression crystallization and preliminary X-ray crystallographic studies of the outer membrane protein OmpW from Escherichia coli. Acta Crystallograph Sect. F. Struct. Biol. Cryst. Commun., 62(4): 415–418.

Baldermann C, Lupas A, Lubieniecki J, Engelhardt H. 1998. The regulated outer membrane protein Omp21 from Comamonas acidovorans is identified as a member of a new family of eight-stranded b-sheet proteins by its sequence and properties. J. Bacteriol., 180(15):3 741–3 749.

Beher M G, Schnaitman C A, Pugsley A P. 1980. Major heatmodifiable outer membrane protein in gram-negative bacteria: comparison with the ompA protein of Escherichia coli. J. Bacteriol., 143(2): 906–913.

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.

Chakrabarti S R, Chaudhuri K, Sen K, Das J. 1996. Porins of Vibrio cholerae: purification and characterization of OmpU. J. Bacteriol., 178(2): 524–530.

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.

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)

Fang H M, Ling K C, Ge R, Sin M. 2000. Enhancement of protective immunity in blue gourami, Trichogaster trichopterus (Pallas), against Aeromonas hydrophila and Vibrio anguillarum by A hydrophila major adhesion. J. Fish Dis., 23: 137–145.

Gordon L A, Alistair J R. 1992. Vaccines, vaccination and the immune response. Lippincott-raven publication, San Diego, California. p.137–143.

Hong H, Patel D R, Tamm L K, van den Berg B. 2006. The outer membrane protein OmpW forms an eight-stranded β-barrel with a hydrophobic channel. J. Biol Chem., 281(11): 7 568–7 577.

Johnson D E, Calia F M, Musher D M. 1984. A Resistance of Vibrio vulnificus to serum bactericidal and opsonizing factors: relation to virulence in suckling mice and humans. J. Infect. Dis., 150(3): 413–418.

Jung C R, Park M J, Heo M S. 2005. Immunization with major outer membrane protein of Vibrio vulnificus elicits protective antibodies in a murine model. J. Microbiol., 43(5): 437–442.

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.

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.

Nandi B, Nandy R K, Mukhopadhyay S, Nair G B, Shimada T, Ghose A C. 2000. Rapid method for species-specific identification of Vibrio cholerae using primers targeted to the gene of outer membrane protein OmpW. J. Clin Microbiol., 38(11): 4 145–4 151.

Nandi B, Nandy R K, Sarkar A, Ghose A C. 2005. Structural features, properties and regulation of the outer-membrane protein W (OmpW) of Vibrio cholerae. Microbiology, 151(9): 2 975–2 986.

Qian R, Chu W, Mao Z, Zhang C, Wei Y, Yu L. 2007. Expression, characterization and immunogenicity of a major outer membrane protein from Vibrio alginolyticus. Acta Biochimica et Biophysica Sinica, 39(3): 194–200.

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.

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

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 L, Wang F, Ye D, Xiao X, Wang S, Peng X. 2006. OmpW and OmpV are required for NaCl regulation in Photobacterium damsela. J. Proteome Res., 5(9): 2 250–2 257.

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.

Xu C, Ren H, Wang S, Peng X. 2004. Proteomic analysis of salt-sensitive outer membrane proteins of Vibrio parahaemolyticus. Res. Microbiol., 155(10): 835–842.

Xu C, Wang S, Ren H, Lin X, Wu L, Peng X. 2005. Proteomic analysis on the expression of outer membrane proteins Vibrio alginolyticus at different sodium concentrations. Proteomics., 5(12): 3 145–3 152.

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.

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