pEGFP/MOMP真核表达重组体的构建及表达  

Construction and sequence analysis of eukaryotic expression recombinant of pEGFP/MOMP

作　　者：粟盛梅[1] 李忠玉[1] 余敏君[1] 占利生[1] 唐双阳[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

出　　处：《中国麻风皮肤病杂志》2005年第7期509-511,共3页China Journal of Leprosy and Skin Diseases

基　　金：湖南省自然科学基金(04JJ3063);湖南省教育厅课题(03C379);湖南省卫生厅课题(B2003-079)

摘　　要：目的:克隆沙眼衣原体主要外膜蛋白(MOMP)基因,构建pEGFP/MOMP真核表达重组质粒,为沙眼衣原体核酸疫苗的研制提供依据。方法:用PCR方法从D型Ct基因组中扩增MOMP全基因片段,克隆入真核表达载体pEGFP相应的酶切位点中,阳性重组子经BamHI、KpnI双酶切、PCR扩增及测序鉴定;并经脂质体介导转染HeLa细胞,36h后观察瞬时表达情况。结果:PCR扩增得到约1.2kb的特异性MOMP基因片段;序列测定证实与GenBank登录的D型沙眼衣原体一致;筛选鉴定出真核表达重组体pEGFP/MOMP。结论:成功地构建了pEGFP/MOMP真核表达重组体,并在HeLa细胞中表达了MOMP。Objective: To clone MOMP gene from genomic DNA of Chlamydia trachomatis and to construct the eukaryotic expression plasmid of pEGFP/MOMP. Methods: MOMP gene was amplified by polymerase chain reaction (PCR) and the fragment was cloned into the vector pEGFP. The positive recombinant was identified by restrictive enzyme digestion, PCR amplification and sequencing. Results: About 1.2kb MOMP gene was successfully isolated. The DNA sequence of MOMP was found to be the same as the nucleotide sequence published by GenBank. pEGFP/MOMP was successfully screened out. Conclusion: pEGFP/MOMP has been constructed successfully and MOMP can be expressed in HeLa cells.

关 键 词：真核表达重组体 PEGFP MOMP 真核表达重组质粒 HeLa细胞 沙眼衣原体 PCR扩增 主要外膜蛋白 真核表达载体 全基因片段 PCR方法 阳性重组子 脂质体介导 核酸疫苗 表达情况 序列测定 基因组 克隆人 Kpn 特异性 D型 酶切 鉴定 

分 类 号：R392[医药卫生—免疫学]