沙眼衣原体主要外膜蛋白在减毒鼠伤寒杆菌中的表达及 …

目的 采用作为疫苗载体的减毒鼠伤寒杆菌致死性平衡系统，构建能异源表达沙眼衣原体（Ｃｔ）主要外膜蛋白（ＭＯＭＰ）的重组菌株。方法 以Ｄ型Ｃｔ的ＤＮＡ为模板，用所设计的特异性引物扩增编码Ｃｔ ＭＯＭＰ高变区（ＶＤＩ～ＶＤＩＶ）基因，并将扩增产物定向克隆至质粒ｐＵＣ１９中。序列分析后，再亚克隆至与减毒鼠伤寒杆菌Ｘ４５５０互补的质粒ｐＹＡ３３４１中，以重组质粒转化减毒鼠伤寒杆菌Ｘ４５５０。结果 对构建的重

Expression and identification of Chlamydia trachomatis major outer membrane protein in attenuated Salmonella typhimurium

Aim To construct a recombinant strain expressing Chlamydia trachomatis major outer membrane protein(MOMP) by using Salmonella typhimurium lethal balance system as vaccine vector. Methods DNA of C. trachomatis serovar D was extracted as template, and a pair of primers flank VS1 to VS4 of MOMP were used to amplify the target gene. The amplified fragment with 780 bp was first cloned into plasmid vector pUC19 and after sequencing was then subcloned into plasmid vector pYA3341, which was a complement of attenuated S. typhimuriumX4550. The later recombinant plasmid was transformed into X4550 by electroporation. Results The recombinant named X4550(pYA3341 780) could express protein of partial MOMP with relative molecular mass(Mr)of 33 000 confirmed by Western blot analysis with species specific monoclonal antibody against MOMP of C. trachomatis. Conclusion MOMP of C. trachomatis was successfully expressed in attenuated S. typhimurium.