摘要
苜蓿银纹夜蛾核型多角体病毒囊膜蛋白Gp64是杆状病毒感染宿主细胞的重要功能蛋白之一.通过Oligo6.0设计一对特异性引物,用于扩增Gp64基因的部分DNA片段,对聚合酶链式反应扩增得到的DNA片段进行纯化,并对纯化后的双酶切DNA片段与经同尾酶双酶切后的原核表达载体p ET28a进行连接;对捕获有重组质粒p ET28a-Gp64的大肠杆菌BL21(DE3)细胞进行异丙基硫代半乳糖苷诱导,通过SDS-PAGE对诱导产物进行电泳分析,结果表明扩增的目的片段获得了表达;采用电透析的方法纯化了该蛋白,为进一步制备抗Gp64蛋白抗体做准备.
Gp64 protein is one of important function proteins in baculovirus virion during infectious process.Specific primers were designed by Oligo6.0 to amplify and purify a truncated fragment of silk worm autographa californica multiple nucleopolyhedro virus Gp64 gene with polymerase chain reaction The purified target DNA fragment was subjected to double enzymatic digestion with Eco RⅠ and XhoⅠ while prokaryotic expression vector p ET28 a was cut by Eco R I and Sal I, then the ligation between the two products was carried out; E.coli BL21（DE3） cells that had caught the recombinant plasmid p ET28a-Gp P64 were induced with isopropyl-β-D-thiogalactoside, the induced product was then analyzed with SDS-PAGE. And the results show that the amplified target is expressed. The protein was purified by electro dialysis, which makes for the preparation of anti Gp64 antibody.
出处
《武汉工程大学学报》
CAS
2015年第7期11-15,共5页
Journal of Wuhan Institute of Technology
基金
湖北省自然科学重点基金(2013CFA101)
