拟态弧菌OmpK基因的克隆及原核表达  

Cloning and Prokaryotic Expression of OmpK Gene of Vibrio mimicus

作　　者：李研东[1] 李菲 卢士英[1] 任洪林[1] 周玉[1] 朱元银 宫彬彬[1] 柳增善[1] 

机构地区：[1]人兽共患病研究教育部重点实验室吉林大学人兽共患病研究所畜牧兽医学院,长春130062 [2]河北省饲料工业协会秘书处,石家庄050031 [3]宁夏中卫畜牧动检站,中卫755000

出　　处：《中国畜牧兽医》2009年第6期52-55,共4页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(30371059)

摘　　要：克隆拟态弧菌(Vibrio mimicus)OmpK(outer membrane protein K)基因并进行相关序列的分析。根据已发表的拟态弧菌OmpK基因序列设计合成引物,以拟态弧菌CGMCC 1.1969标准株的基因组DNA为模板进行PCR扩增,获得801bp片段并测序,序列已提交至GenBank(GenBank登录号为FJ462707)。拟态弧菌OmpK序列比对结果显示,溶藻弧菌、副溶血弧菌、创伤弧菌、河弧菌OmpK基因序列的同源性均大于78%。该基因编码1个含267个氨基酸残基的蛋白质,预测分子质量为29.6 ku。利用基因工程方法将该基因片段定向克隆至表达载体中,并在大肠杆菌中获得高效表达,表达后的OmpK蛋白分子质量约为28 ku。A target of common antigen, OmpK （outer membrane protein K）, is the receptor for KVP40 which is abroad- host-rang Vibriophage. To investigate the possibility of OmpK as species antigen, primers were designed according to OmpK gene sequence published in GenBank, and a piece of DNA sequence 801 bp was amplified by PCR from genomie DNA of Vibrio mirnicus. The OmpK gene of Vibrio mimicus was cloned with 801 nucleotides encoding 267 amino acids. The OmpK gene was expressed in Escherichia coli induced by IPTG. The molecular weight of recombinant protein was about 28 ku.

关 键 词：拟态弧菌 OmpK基因 克隆 表达 

分 类 号：Q78[生物学—分子生物学]