摘要
目的构建梅毒螺旋体TpN15-47融合基因及其原核表达系统。方法分别克隆TpN15和TpN47基因,利用T4连接酶构建TpN15-47融合基因,常规方法构建其原核表达系统。采用SDS-PAGE检测目的重组蛋白TpN15-47表达情况。结果连接的TpN15-47基因测序分析表明,插入的基因片段为TpN15-47融合基因,与GenBank报道相同。目的重组蛋白TpN15-47表达产量约为细菌总蛋白85.79%,主要以包涵体形式存在。结论所构建的原核表达系统能高效地表达TpN15-47融合蛋白,为后期梅毒螺旋体基因疫苗和临床检测试剂盒的研制奠定了基础。
Objective To construct TpN15-47 fusion gene and its prokaryotic expression system.Methods The gene of TpN15 and TpN47 were cloned respectively by PCR, TpN15-47 fusion gene was constructed using T4 ligase and prokaryotic expression of the fusion gene done with routine techniques. SDS-PAGE was used to examine expression of the target recombinant protein TpN15-47. Resuits The lined gene＇s sequence TpN15-47 DNA was identical with that gotten by dealed with the dates from the GenBank.Expression output of the target recombinant protein TpN15-47, mainly present in inclusion body, accounted for 85.79% of the total bacterial proteins. Condusion The prokaryotic expression systems with high efficiency of TpN15-47 fusion gene was successfully constructed, which may help in the further research of genetic engineering vaccine and clinical detection of Treponema pallidum.
出处
《中国实验诊断学》
2007年第12期1661-1663,共3页
Chinese Journal of Laboratory Diagnosis
