摘要
从真核表达载体pEGFP-NS2中克隆得到猪瘟病毒(CSFV)NS2 3′端的基因片段,将其克隆到pET-32a载体上,构建出重组表达载体,经IPTG诱导表达。结果表明,克隆了大小为363 bp NS2 3′端的基因,重组载体pET-NS2-C363在大肠埃希菌中以包涵体形式表达,表达产物经SDS-PAGE和Western blot鉴定大小为33.84 ku,与预期结果一致。结果为进一步研究NS2蛋白与猪脐静脉血管内皮细胞中蛋白之间的相互作用提供材料及为制备单克隆抗体奠定基础。
The research studied cloning and expression of NS2 gene.363 bp of NS2 gene of CSFV Shimen strain was amplified by PCR,and cloned into prokaryotic expression vector pET-32a.The target protein was expressed in E.coli BL21(DE3) induced with IPTG,and detected by SDS-PAGE and Western blot.The results indicated that the 33.84 ku expressed protein was obtained.,It laid the foundation to study the interaction between theporcine vascular endothelia cell protein andNS2 protein and monoclonal antibody preparatio.
出处
《动物医学进展》
CSCD
北大核心
2011年第5期14-17,共4页
Progress In Veterinary Medicine
基金
国家自然科学基金项目(30972186)
关键词
猪瘟病毒
NS2基因
克隆
原核表达
Classical swine fever virus
NS2 gene
coling
prokaryotic expression