沙眼衣原体D型MOMP基因原核表达载体的构建及序列分析

Construction of a prokaryotic expresssion vector carrying MOMP gene from chlamydia trachomatis serovar D and partial sequence determination

目的:构建沙眼衣原体D型MOMP基因的原核表达载体,为沙眼衣原体基因工程疫苗的研究提供材料。方法:PCR技术扩增D型沙眼衣原体标准株的MOMP基因片段,将其定向插入原核表达载体pRSET多克隆位点中,构建重组表达载体。并用酶切分析、PCR扩增及序列测定方法对重组载体进行鉴定。结果:沙眼衣原体MOMP基因被正确克隆到原核表达载体pRSET-A中,测序结果与Genebank中AF279589等菌株序列有99.99%同源性。结论:原核表达载体pRSET-MOMP构建成功。

Objeetive： To construct a prokaryotic expression vector carrying chlamydia trachomatis major outer membrane protein （MOMP） gene. Methods：The MOMP gene was amplified from the genome of the chlamydia trachomatis by PCR and recombined with plasmid pRSET - A. The recombinant plasmid pRSET - MOMP was verified with restriction analysis, PCR and sequence. Results： MOMP gene was cloned correctly into vector pRSET - A. Conclusion： The prokaryotic expression vector pRSET - MOMP is constructed successfully.