沙眼衣原体假定蛋白CT389原核表达质粒的构建及结构预测  

Construct the prokaryotic expression plasmid encode hypothetical protein CT389 from Chlamydia trachomatis and structure prediction

作　　者：唐翠连[1,2] 陈浩[1] 刘玖林 周洲[1] 

机构地区：[1]南华大学医学院病原生物学研究所,湖南衡阳421001 [2]湖南省邵阳学院附属医院检验科 [3]南华大学医学院

出　　处：《中南医学科学杂志》2017年第4期331-334,共4页Medical Science Journal of Central South China

基　　金：国家自然科学基金项目(No.31570179;30901352);湖南省自然科学基金(No.13JJ4072);湖南省教育厅优秀青年基金(No.12B109);湖南省分子靶标新药研究协同创新中心资助项目(No.2015-351);特殊病原体防控湖南省重点实验室资助项目(No.2014-5号);湖南省重点学科资助项目(No.2011-76)

摘　　要：目的构建能表达沙眼衣原体(Chlamydia trachomatis,Ct)假定蛋白CT389的质粒pGEX6P-ct389及预测其结构。方法设计特异性引物,以Ct基因组为模板,PCR法扩增ct389基因,构建携带目的基因的重组质粒pGEX6Pct389;通过PCR鉴定和双酶切鉴定初筛,测序鉴定验证结果;比较ct389和tc0668基因序列及其编码产物的相似性,采用生物信息学预测CT389蛋白的二级结构和三级结构。结果获得大小约为1 300 bp的ct389基因片段,并成功得到重组质粒pGEX6P-ct389。ct389和tc0668基因及其编码产物的相似性高达84%和92%,CT389蛋白α-螺旋、无规卷曲和β片层的含量分别为27.7%、49.26%和占23.04%。与其三级结构最接近的是鼠疫耶尔森氏菌的纤溶酶原激活物。结论Ct389与tc0668基因从DNA到编码产物具有较高的相似性,ct389可能是重要的沙眼衣原体毒力基因。Objective To construct the prokaryotic expression plasmid pGEX6P-ct389 which express Chlamydia trachomatis hypothetical protein CT389 and predict its protein structure.MethodThe specific primers were designed and the Ct 389 gene was amplified through PCR by using Ct genome as template to construct prokaryotic expression plasmid pGEX6P-ct389 which contain the ct389 gene.The similarity between ct389 and tc0668 gene and their proteins were compared.The binary and ternary structure were analyzed by bioinformatics.ResultsThe 1300 bp length of Ct389 gene was gained and the recombinant plasmid pGEX6P-Ct389 was constructed successfully.The similarities of ct389 and tc0668 genes and their coding products were up to 84% and 92% respectively.The content of alpha-helix,random coil and beta sheet of CT389 protein are 27.7%,49.26% and 23.04%.The ternary structure of CT389 protein is similar to the structure of Pla protein from Yersinia pestis.ConclusionFrom DNA to coding product,ct389 has high similarities with tc0668 gene,and ct389 gene may be an important Chlamydia trachomatis virulence factor.

关 键 词：沙眼衣原体 蛋白结构 假定蛋白 CT389 

分 类 号：R374.1[医药卫生—病原生物学]