Expression and purification of two major outer membrane proteins from <Emphasis Type="Italic">Vibrio alginolyticus</Emphasis> |
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Authors: | Ronghua Qian Zhaohua Xiao Chongwen Zhang Wuying Chu Zhijuan Mao Lian Yu |
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Institution: | (1) Department of Microbiology, College of Life Sciences, Hunan Normal University, Changsha, 410081, China;(2) Institute of Preventive Veterinary Medicine, Zhejiang University, Hangzhou, 310029, China;(3) Xiangya Hospital of Central South University, Changsha, 410008, China;(4) Institute of Subtropical Agriculture, The Chinese Academy of Sciences, Changsha, 410125, China |
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Abstract: | Vibrio alginolyticus, a Gram-negative bacterium, is one of Vibrio pathogens common to human and marine animals. Outer membrane proteins of bacteria play an important role during infection
and induction of host immune response. In present research, two outer membrane protein genes (OmpK and OmpW) of V. alginolyticus were cloned and expressed. The open reading frames of OmpK and OmpW contain 846 bp and 645 bp, respectively, the mature proteins consist of 261 and 193 amino acid residues. At the signal peptides
positions −3 to −1, the amino acids were V-M-A in OmpK and V-F-A in OmpW, which consistented with the observed sequence V-X-A of the signal peptides of transmembrane OMP. The alignment analysis
indicated that both proteins were highly conserved, which could serve as surface antigens for vaccine candidates. SDS-PAGE
indicated two genes over-expressed in E. coli BL21 (DE3). By affinity chromatography on Ni2+-nitriloaceate resin, the recombinant proteins were purified from inclusion bodies. Western blot analysis revealed that both
proteins had immunoreactivity, which provided a base for further study on the evaluation of diagnostication and vaccine candidates. |
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Keywords: | Characterization Expression Outer membrane proteins Purification Vibrio alginolyticus |
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