新烟草糖基转移酶原核表达质粒的构建及诱导

Construction and Inducement of the Prokaryotic Expressed Plasmid of New Glycosyltransferases from Tobacco

利用先期从烟草中克隆的一个糖基转移酶基因,通过PCR扩增得到该基因的ORF,将此片段与原核表达载体pTYB1连接,构建重组载体pT,然后转入大肠杆菌,并进行诱导表达,SDS-PAGE检测表达产物。结果表明,此基因的ORF可在大肠杆菌中表达,但表达产物以包涵体形式存在。

The ORF of a new tobacco glycosyltransferases gene cloned by the lab early was amplified through PCR. Then this fragment was cloned into the prokaryotie expression pTYBI to generate the recombinant plasmid pT. After the recombinant pT expressed by IPTG inducing in Escherichia coli strain ER2566, the crude cell extract was analyzed by SDS-PAGE. The result showed that the tobacco glycosyltransferases fusion protein has been expressed successfully, but it existed as inclusion body. insoluble form.