溶藻弧菌HY9901鞭毛蛋白flaC基因的克隆和序列分析及真核表达质粒的构建

journal6 ›› 2010, Vol. 31 ›› Issue (3): 95-100.

溶藻弧菌HY9901鞭毛蛋白flaC基因的克隆和序列分析及真核表达质粒的构建

（1.广东海洋大学水产学院，广东湛江524088；2.广东省水产经济动物病原生物学及流行病学重点实验室，广东湛江524088;3.广东省水产经济动物病害控制重点实验室，广东湛江524088）

出版日期:2010-05-25 发布日期:2012-04-17
通讯作者: 吴灶和（1953-），男，广东揭阳人，广东海洋大学教授，博士，主要从事海洋生物学研究；E-mail：wuzh@gdou.edu.cn
作者简介:梁海鹰（1971-），女，湖南怀化人，广东海洋大学水产学院副教授，博士，主要从事海洋生物学研究
基金资助:
国家科技支撑计划项目（2007BAD29B03）；广东省自然科学基金项目（9151064201000063）；广东海洋大学自然科学研究项目（0612187）

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

(1.Fisheries College,Guangdong Ocean University,Zhanjiang 524088,Guangdong China;2.Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals,Zhanjiang 534088,Guangdong China3.Guangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals,Zhanjiang 524088,Guangdong China)

Online:2010-05-25 Published:2012-04-17

摘要/Abstract

摘要：参照GenBank上登录的副溶血弧菌鞭毛蛋白flaC基因序列设计引物，PCR扩增溶藻弧菌HY9901株的flaC全长基因，序列分析结果显示该基因为1 155 bp，编码384个氨基酸.与GenBank中其他弧菌的同源基因序列比对显示，溶藻弧菌flaC基因与副溶血弧菌flaC基因的同源性最高(87%).将该基因定向克隆到真核表达质粒pcDNA3.1(+)中，获得带溶藻弧菌鞭毛蛋白flaC基因的真核表达重组质粒pcDNA-flaC，为其DNA疫苗的进一步研究奠定了基础.

关键词： 溶藻弧菌, flaC基因, 基因克隆, 序列分析, 重组真核表达质粒

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

梁海鹰, 夏立群, 吴灶和, 简纪常. 溶藻弧菌HY9901鞭毛蛋白flaC基因的克隆和序列分析及真核表达质粒的构建[J]. journal6, 2010, 31(3): 95-100.

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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