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机构地区:[1]深圳市赛尔生物技术有限公司,深圳518055 [2]扬州大学医学院病原学与免疫学教研室 [3]深圳市人民医院检验科
出 处:《现代预防医学》2011年第3期529-532,共4页Modern Preventive Medicine
摘 要:[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] To construct and identify the procaryotic expression vector of the powerful epitopes of gG-1. [Methods] The domains of the powerful epitopes were analyzed by DNAstar and amplified by PCR, then the production was directly cloned into procaryotic expression vector pGEX-4T-1 for construction of pGEX-4T-1/gG-1, the recombinant plasmid was identified by restriction endonuclease and DNA sequencing. [Results] The recombinant plasmid was successfully constructed, the comparison of inserted fragment with that reported in GenBank revealed that the identification was very high and the inserted fragment was in proper order. [Conclusion] The successful construction of pGEX-4T-1/gG-1 lays a good foundation for the research of new type-specific serological detection of HSV-1.
分 类 号:R752.11[医药卫生—皮肤病学与性病学]
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