摘要
以质粒pGEM-pvs为模板,经PCR扩增得到了长约890bp的PVS-CP基因条带。回收特异性DNA带并与T表达载体pBAD/Thio-TOPO连接,连接物转化受体菌获得了Amp抗性菌落。经PstⅠ、SphⅠ酶切鉴定,筛选到了PVS-CP基因正向插入载体的阳性克隆。DNA测序结果表明PVS-CP基因插入方向与读码正确,成功构建了PVS-CP基因的原核表达载体。经阿拉伯糖诱导获得了预期的49kDa融合蛋白,PVS-CP基因在受体菌中表达正确。
Using pGEM -pvs as template, PCR -amplified potato visus S (PVS) coat protein (CP) gene fragment about 890bp was obtained. The specific DNA fragment was recovered and ligated into the expression vector pBAD/ Thio- TOPO. Pst Ⅰand Sph Ⅰ restriction analysis indicated that PVS - CP gene was inserted into the expression vector in direct orientation. DNA sequencing confirmed that PVS - CP gene was inserted into the vector in direct orientation and in frame. After induction of arabinose, specific 49kDa fusion protein was found, PVS - CP gene was expressed correctly in E. coli.
出处
《青岛农业大学学报(自然科学版)》
2007年第3期159-161,共3页
Journal of Qingdao Agricultural University(Natural Science)
基金
青岛市自然科学基金项目(05-1-JC-910)资助
关键词
马铃薯S病毒
CP基因
原核表达载体
potato virus S
coat protein gene
prokaryotic expression vector