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[目的]构建gG-1强抗原决定簇集中区对应基因的重组原核表达载体,并对其进行鉴定。[方法]利用DNAstar软件分析gG-1强抗原决定簇的集中区,PCR扩增相应基因片段,将其定向克隆至原核表达载体pGEX-4T-1中,构建重组质粒pGEX-4T-1/gG-1,并对其进行双酶切和DNA测序鉴定。[结果]成功构建了pGEX-4T-1/gG-1重组原核表达载体,且目的基因片段与GenBank中已公布序列一致性较高,阅读框架正确。[结论]pGEX-4T-1/gG-1重组原核表达载体的成功构建,为研制HSV-1型特异性基因工程诊断试剂奠定了基础。
[Objective] The aim of the study was to construct a recombinant prokaryotic expression vector containing the corresponding gene of gG-1 strong antigenic determinant and to identify it. [Method] The DNA gG-1 gene fragment was amplified by DNAstar software and cloned into the prokaryotic expression vector pGEX-4T-1. The recombinant plasmid pGEX-4T-1 / gG -1, and its double digestion and DNA sequencing identification. [Result] The recombinant prokaryotic expression vector pGEX-4T-1 / gG-1 was constructed successfully. The sequence of the target gene was consistent with the published sequences in GenBank and the reading frame was correct. [Conclusion] The successful construction of pGEX-4T-1 / gG-1 recombinant prokaryotic expression vector lays a foundation for the development of HSV-1 specific genetic engineering diagnostic reagent.