HIV-1 Tat基因在pEGx-KG中的融合构建及原核表达

目的:在大肠杆菌BL21(DE3)中高效表达融合的Tat蛋白。方法:用PCR方法从HIV cDNA文库中扩增出Tat基因,并将其插入到表达载体pEGx-KG中,转化到E.coli中进行融合表达,表达产物用蛋白电泳和Western-blot进行鉴定。结果:成功地构建了重组质粒pEGx-KG-Tat,重组质粒在大肠杆菌BL21(DE3)中得到高效表达。蛋白电泳和Western-blot分析表明,在相对分子质量(Mr)为38.9kDa处有1条特异性的带。结论:GST基因与HIV-1 Tat基因的融合构建,使Tat蛋白在大肠杆菌中得到高效表达,为艾滋病的防治研究奠定了基础。

The fusion construction of HIV-1 Tat gene in pEGx-KG and efficient expression in Escherichia coli

Objective: To study the efficient expression of GST-Tat protein in Escherichia coli BZ21(DE3).Methods: HIV-1 Tat gene was amplified by PCR from cDNA library of HIV and was inserted into vector of pEGx-KG.The recombinant plasmid was transferred and expressed in E.coli BL21(DE3).The expressed products were identified by SDS-page and Western-blot.Results: HIV-1 Tat gene was amplified successfully by PCR.The recombinant plasmid was expressed efficiently in E.coli(BL21).SDS-page and Western-blot analyses showed the expressed Tat fusion protein with relative molecular weight was 38.9 kDa.Conclusion: HIV-1 Tat gene can be cloned and GST-Tat fusion protein can be expressed efficiently in E.coli,which may contribute to further research of anti-AIDS.