›› 2009, Vol. 36 ›› Issue (6): 52-55.

• 生物技术 • 上一篇    下一篇

拟态弧菌OmpK基因的克隆及原核表达

李研东1,李菲2,卢士英1,任洪林1,周玉1,朱元银3,宫彬彬1,柳增善1   

  1. (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)
  • 收稿日期:1900-01-01 修回日期:2009-03-24 出版日期:2009-06-20 发布日期:2009-06-20
  • 通讯作者: 柳增善

Cloning and Prokaryotic Expression of OmpK Gene of Vibrio mimicus

LI Yan-dong1,LI Fei2, LU Shi-ying1,REN Hong-lin1,ZHOU Yu1, ZHU Yuan-yin3, GONG Bin-bin1,LIU Zeng-shan1   

  1. (1.Key Laboratory for Zoonosis Research, Ministry of Education, Institute of Zoonosis,College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China; 2.Hebei Feed Industry Association, Shijiazhuang 050031, China; 3.Animal Quarantine Station of Ningxia Zhongwei, Zhongwei 755000, China)
  • Received:1900-01-01 Revised:2009-03-24 Online:2009-06-20 Published:2009-06-20

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801 bp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。

关键词: 拟态弧菌; OmpK基因; 克隆; 表达

Abstract: 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 genomic DNA of Vibrio mimicus. 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.

Key words: Vibrio mimicus; OmpK gene; clone; expression

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