Asia1型口蹄疫病毒VP1基因的克隆、原核表达及纯化

口蹄疫(Foot-and-mouthdisease，FMD)是由口蹄疫病毒感染引起的偶蹄动物共患的急性、热性、接触性传染病，该病被国际兽疫局列为A类传染病之首，对畜牧业危害极大，是世界各国检疫和防疫的重点对象。目前已知该病毒有7个血清型，

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

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.