Cloning and Sequencing of flaC Gene of Vibrio Alginolyticus Strain HY9901 and Construction of Its Eukaryotic Expression Recombinant Plasmid

Abstract

Abstract: To investigate the possibility of flaC as a candidate antigen for vaccine production,primers were designed based on flaC gene sequences published in GenBank.The flaC gene of Vibrio alginolyticus HY9901 was amplified by PCR and cloned into pMD19-T vector.Sequence analysis revealed that flaC gene is 1 155 bp and encodes a putative protein of 384 amino acids.The flaC gene sequence of V. algonilyticus showed highest identity to V. parahaemolytus (87%).The PCR product was cloned into eukaryotic expression vector pcDNA3.1(+) and the positive clone was chosen and identified through enzyme digestion analysis and sequencing.An eukaryotic expression recombinant plasmid,pcDNA-flaC,containing the flaC gene of flagellin in V. alginolyticus was constructed,which would be a foundation for further study on its DNA vaccine.

Key words: vibrio alginolyticus, flaC gene, gene clone, sequence analysis, eukaryotic expression recombinant plasmid

LIANG Hai-Ying, XIA Li-Qun, WU Zao-He, JIAN Ji-Chang. Cloning and Sequencing of flaC Gene of Vibrio Alginolyticus Strain HY9901 and Construction of Its Eukaryotic Expression Recombinant Plasmid[J]. journal6, 2010, 31(3): 95-100.

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