摘要
利用PCR技术扩增出水貂阿留申病毒(ADV)含有VP2抗原表位区的基因片段,将其克隆到原核表达载体pGEX-4T-1中,构建出重组质粒pGEX-4T-VP2,并转化入大肠杆菌BL21中,用IPTG法进行诱导表达。经SDS-PAGE凝胶电泳和免疫印迹分析显示有目的条带,并且在诱导6 h后表达量达到最高,随时间的延长表达量降低。本研究成功构建了pGEX-4T-VP2重组质粒,确定了VP2基因的优化表达条件,为阿留申病的免疫学诊断和疫苗研制奠定基础。
A gene fragment with VP2 epitope of Aleutian mink disease virus（ADV） was amplified by PCR and was cloned into the prokaryotic expression vector pGEX-4T-1,constructing the recombinant plasmid pGEX-4T-1-VP2.The later was transformed into E.coli BL21,induced to express with IPTG.The target band was found by SDS-PAGE gel electrophoresis and western blotting analysis.The expressed reached the highest level at and reduced after 6 hours.The recombinant plasmid pGEX-4T-VP2 was successfully constructed in the study and the optimal expression conditions of VP2 gene was determined.The information provided by the study is useful for the immunological diagnosis and vaccine development of Aleutian disease.
出处
《经济动物学报》
CAS
2013年第1期12-14,18,共4页
Journal of Economic Animal
关键词
水貂阿留申病毒
VP2
原核表达
Aleutian mink disease parvovirus
VP2 gene
prokaryotic expression
