西尼罗病毒NS1基因的原核表达与抗原性鉴定  

Prokaryotic expression of West Nile virus NS1 gene

作　　者：马健男[1] 杨涛[1] 王群[1] 张维军[1] 徐青元[1] 田野[1] 林燕[1] 宋红梅[1] 步志高[1] 吴东来[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/农业部兽医公共卫生重点开放实验室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2009年第8期592-595,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家863计划(No.2007AA02Z406)

摘　　要：为进行西尼罗病毒(WNV)NY99株NS1蛋白的免疫学研究,通过PCR方法扩增WNV NY99株NS1基因,并将其克隆至原核表达载体pMAL-c2X上,构建出重组表达质粒pc2X-NS1,经IPTG诱导得到可溶性的麦芽糖结合蛋白(MBP)-NS1融合蛋白(MBP-NS1),其分子质量约为90ku,约占可溶性菌体蛋白质量的50%。利用直链淀粉树脂柱对MBP-NS1纯化后的重组蛋白的纯度达到60%,western blot和间接ELISA检测证明纯化的MBP-NS1具有良好的抗原性和特异性,为进一步开展关于WNV NS1蛋白的研究奠定了基础。The NS1 protein of West Nile virus NY99 strain was amplified by PCR with specific primers and cloned into pMAL-c2X. The recombinant vector was transformed into E. coli TB1 competent cells and a fusion protein MBP-NS1 was expressed after induction with IPTG. The MBP-NS1 protein was purified and analysed by western blot and indirect enzyme-linked immunosorbent assay. The results showed that the purified fusion protein retained good antigenicity and specificity.

关 键 词：西尼罗病毒 NS1基因 原核表达 

分 类 号：S852.61[农业科学—基础兽医学]