轮状病毒VP4蛋白与热稳定肠毒素融合基因的克隆及其植物表达载体的构建

Cloning of VP4-ST gene fused from rotavirus VP4 protein gene with heat-stable toxin gene and construction of recombinant plant expression plasmid pB-VP4-ST

利用DNA重组技术,将用PCR扩增来的 VP4-ST融合基因分别克隆到原核表达载体pThioHisB及植物表达载体 pBin438 中,成功构建了重组表达质粒 pTh-VP4-ST和 pB-VP4-ST。将pTh-VP4-ST进行SDS-AGE分析,结果表明,表达的目的蛋白以包涵体形式存在,大小约 40ku;同时将pB-VP4-ST转化了农杆菌EHA105。

The recombinant expression plasmids (pTh)-VP4-ST and pB-VP4-ST were successfully constructed by cloning the VP4-ST fused gene fragments, which were amplified with a pair of primers by PCR, into a prokaryotic expression vector pThioHis and a plant expression vector pBin438, respectively. The fusion protein VP4-ST was expressed from the prokaryotic expression plasmid pTh-VP4-ST in BL21 by inducing with IPTG. The SDS-PAGE analysis showed that the expressed fusion protein is 40 ku in size. In addition, the plant expression plasmid pB-VP4-ST was transformed into the Agrobacterium EHA105.