Gene Cloning and Expression of Outer Membrane Protein OmpU of Vibrio vulnificus

LI Su-yi; WU Wei-wei; LI Pan; KE Ling; XU Bin-fu; LIN Chen-tao; LIN Tian-long; CHEN Xu

doi:10.19303/j.issn.1008-0384.2013.06.001

Volume 28 Issue 6

Jun. 2013

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LI Su-yi, WU Wei-wei, LI Pan, KE Ling, XU Bin-fu, LIN Chen-tao, LIN Tian-long, CHEN Xu. Gene Cloning and Expression of Outer Membrane Protein OmpU of Vibrio vulnificus[J]. Fujian Journal of Agricultural Sciences, 2013, 28(6): 517-521. doi: 10.19303/j.issn.1008-0384.2013.06.001

Citation:

LI Su-yi, WU Wei-wei, LI Pan, KE Ling, XU Bin-fu, LIN Chen-tao, LIN Tian-long, CHEN Xu. Gene Cloning and Expression of Outer Membrane Protein OmpU of Vibrio vulnificus[J]. Fujian Journal of Agricultural Sciences, 2013, 28(6): 517-521. doi: 10.19303/j.issn.1008-0384.2013.06.001

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Gene Cloning and Expression of Outer Membrane Protein OmpU of Vibrio vulnificus

doi: 10.19303/j.issn.1008-0384.2013.06.001

1. Biotechnology Institute,Fujian Academy of Agricultural Sciences,Fuzhou,Fujian 350003,China;
2. Fujian Normal University,Fuzhou,Fujian 350007,China;
3. Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China

Received Date: 2013-05-01
Publish Date: 2013-06-18

Abstract

Abstract

Vibrio vulnificus is an important pathogen that causes the eel sepsis.In this study,the outer membrane protein U(ompU)gene of Vibrio vulnificus strain FJ03-X2is cloned,and then inserted into the prokaryotic expression vector,named pET32a-ompU.The plasmid was then transformed into E.coli strain to induce the expression of rompU protein.The soluble rompU protein can be expressed under the induction of IPTG(0.5mmol/ L)at 37℃for 4h.Then purified it by Ni-NTA His Binding affinity purification.SDS-PAGE displayed the purified rompU as a single band of the expected size.The purified rompU protein was then injected into SD level rats to produce polyclonal antibody.High specificity antiserum of FJ03-X2rompU is confirmed by ELISA detection.Western-blot confirmed that OmpU polyclonal antibodies can recognize the natural outer membrane protein OmpU in Vibrio vulnificus FJ03-X2.
- Vibrio vulnificus,
- outer membrane protein OmpU,
- cloning,
- prokaryotic expression,
- protein purification,
- polyclonal antibody

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References(15)

References

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