摘要
根据AEV-NH937基因组全序列设计3对引物,应用RT-PCR方法,扩增出AEV的外壳蛋白VP1、VP3、VP0基因,将它们克隆于pUCm-T载体上进行序列分析。结果显示,AEV-NH937病毒株VP1基因全长810 bp、编码270个氨基酸,VP3基因全长735 bp、编码245个氨基酸,VP0基因全长726 bp、编码242个氨基酸;与Calnek疫苗株相比,AEV-NH937病毒株VP1、VP3、VP0基因的核苷酸同源性分别为90.62%、93.88%、95.45%,氨基酸同源性分别为88.89%、97.96%、98.35%。将VP1、VP3、VP0基因分别插入载体pcDNA4/His Max构建表达重组质粒并转入宿主菌DH5α中,使基因得以表达。
Three pairs of primer were designed according to avian encephalomyelitis virus（AEV） isolate NH937.The structural protein VP1,VP3 and VP0 genes of AEV were amplified by RT-PCR.These genes were cloned into the plasmid pUCm-T,and the inserts were sequenced.The result of nucleotide sequencing showed VP1,VP3 and VP0 genes consisted of 810 bp,735 bp,726 bp and encoded 270,245,242 amino acids,respectively.The VP1,VP3 and VP0 genes of AEV-NH937 strain shared respectively 90.62%,93.88%,95.45% homology with those from the Calnek vaccine strain of AEV,and that deduced amino acids sequences from the VP1,VP3,VP0 genes of AEV-NH937 strain shared respectively 88.89%,97.96%,98.35% homology with those from the Calnek vaccine strain of AEV.To express VP1,VP3,VP0 in E.coli,three recombinant expression vector were constructed and transformed into E.coli,then incubated at 37℃.
出处
《化学与生物工程》
CAS
2009年第12期70-72,共3页
Chemistry & Bioengineering
基金
内蒙古自治区高等学校科学研究资助项目(NJ03017)
