畜牧兽医学报 ›› 2005, Vol. 36 ›› Issue (6): 631-634.doi:

• 研究简报 • 上一篇    下一篇

Asia1型口蹄疫病毒VP1基因的克隆、原核表达及纯化

独军政;常惠芸; 丛国正;林 彤;邵军军;魏小娟;刘在新;谢庆阁   

  1. 中国农业科学院兰州兽医研究所国家口蹄疫参考实验室, 兰州 730046
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2005-06-25 发布日期:2005-06-25

Gene Cloning,Prokaryotic Expression and Purification of the VP1 of Foot-and-mouth Disease Virus Serotype Asia 1

DU Jun-zheng; CHANG Hui-yun; CONG Guo-zheng;LIN Tong; SHAO Jun-jun;
WEI Xiao-juan;LIU Zai-xin; XIE Qing-ge   

  1. National FootandMouth Disease Reference Laboratory,Lanzhou Veterinary Research Insititute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2005-06-25 Published:2005-06-25

Abstract: According to the published nucleotide sequences of the VP1 gene of footandmouth disease virus serotype Asia 1 isolates, a pair of primers were designed and synthesized to clone the VP1 gene of YNBS/58 strain. PCR product was cloned into pProexHTb vector, and E.coli BL21 was transformed by the recombinant plasmid pProex-VP1 for sequencing and expression .The expressed product was identified by SDS-PAGE and Western blot, and purified by Ni-NTA His.Bind resins. The results showed that the nucleotide sequence identity of VP1 gene between YNBS/58 and India93, India 97,India99, Iseral and YNAs11 strains is from 80.3% to 97.5% and amino acid identity is from 85.8% to 96.5%, the recombinant VP1 protein was significant at 34 ku by SDS-PAGE and Western blot, which accounted for 30% of total protein in E.coli lysates,and the recombinant protein was purified successfully.