风疹病毒E1特异肽段的重组质粒载体构建

Construction of Plasmid Expression Vector for Specific Peptide of the Rubella Virus E1 Gene

目的:构建风疹病毒(RV)E1特异肽段的重组质粒载体,以期表达RVE1特异肽段的重组蛋白。方法:抽提RV减毒活疫苗Wistar RA27/3株的基因组RNA后逆转录,用PCR方法扩增E1特异肽段的基因片段;扩增产物经纯化后克隆至pGEX-2T质粒载体,挑取重组质粒阳性克隆进行双酶切鉴定和测序。结果:成功克隆出330bp左右的目的片段,重组质粒经测序后与预期序列一致。结论:RVE1特异肽段的基因片段的成功克隆及重组质粒的构建为进一步表达相应重组蛋白奠定了基础。

Objective： To construct a recombinant plasmid vector of the RV specific fragment for expressing the specific fragment of RV E1 protein. Methods ： RNA of the RV attenuated live vaccine Wistar RA27 / 3 strain was extracted and reversely transcribed. The specific fragment of the E1 gene was amplified and the PCR products cloned in the vector pGEX-2T after purification. Positive clones were selected and identified by two-enzyme digestion and sequence analysis. Results ： A 330 bp target fragment was successfully cloned, and the sequence of the recombinant plasmid was consistent with the original sequence. Conclusion ： Successful cloning of the RV E1 specific fragment and the construction of the recombinant plasmid have laid a foundation for further expressing the recombinant protein.