Biotechnology Bulletin ›› 2013, Vol. 0 ›› Issue (4): 116-122.

• Research Report • Previous Articles     Next Articles

Molecular Cloning and Bioinformatics Analysis of Outer Membrane Protein H Gene from Vibrio alginolyticus Strain HY9901

Zhou Zejun1，2，3, Pang Huanying1，2，3, Ding Yu1，2，3, Jian Jichang1，2，3, Wu Zaohe2，3，4   

  1. 1. Fisheries College of Guangdong Ocean University，Zhanjiang 524088 ；2. Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals，Zhanjiang 524088 ；3. Key Laboratory of Diseases Controlling for Aquatic Economic Animals of Guangdong Higher Education Institutions，Zhanjiang 524088 ；4. Zhongkai University of Agriculture and Engineering，Guangzhou 510225
  • Received:2012-10-08 Revised:2013-04-22 Online:2013-04-22 Published:2013-04-22

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Abstract: Primers for PCR cloning were designed according to the whole genome sequence of vibrio alginolyticus published in GenBank. The outer membrane protein H（OmpH）gene of V.alginolyticus strain HY9901 was amplified by PCR and cloned into pMD18-T vector to investigate the possibility of OmpH as a candidate antigen for vaccine production. Sequence analysis revealed that OmpH gene（GenBank Number ：JX855924）is 573 bp and encodes a putative protein of 190 amino acids. The predicted molecular weight（MW）of OmpH was 21.1 kD with an estimated pI of 9.02. Using SignalP 4.0 and TMHMM Server 2.0 software, and it was predicted that the OmpH protein did not contain a signal peptide or a transmembranous region. This protein had one N-glycosylation site, six phosphorylation sites, one endoplasmic reticulum targeting sequence and three microbodies C-terminal targeting signals predicted by SoftBerry-Psite software. To further analyze the evolutionary relationship among OmpH, a molecular phylogenetic tree was constructed using MEGA5.0 software. In this tree, the OmpH protein showed high genetic relationship with Vibrio parahaemolyticus and Vibrio harveyi. Using bioinformatices softwares and methods, the B-cell preponderant epitopes of OmpH were localized in the regions of 5-10、106-110、120-125、158-163 and 175-180. The three-dimensional structure of OmpH was determined using SWISS-MODEL work-space and it had a similar structure with Skp protein of Escherichia coli. These results can provide a basis for further studies on the immunogenecity of OmpH and vaccine preparation.