摘要
目的构建表达肺炎衣原体膜表面蛋白(OMP)的重组质粒,并在大肠杆菌中表达获得基因重组蛋白.方法用高保真PCR方法从肺炎衣原体扩增OMP片段,双酶切后插入原核表达质粒pQE 30,在大肠杆菌M15中表达.结果 重组质粒经双酶切鉴定与目的基因长度相符;各表达蛋白经SDS PAGE分析,相对分子量与文献相符.结论 该重组质粒可用于核酸疫苗的备选质粒,基因重组菌表达的融合蛋白有可能作为有效抗原用于肺炎衣原体检测试剂盒的制备.
Objective Constructing the recombinant plasmid of Chlamydia pneumoniae outer membrane protein and expressing fusion protein in E.coli M15. Methods Gene encoding OMP was amplified from C.pn chromosomal DNA by high fidelity PCR. The gene was inserted into the prokaryotic expression vector pQE-30. E.coli M15 was used to express recombinant OMP. Results The length of recombinant DNA was same with PCR products by endonuclease digestion analysis. The calculated molecular mass of fusion protein was confirmed with SDS-PAGE analysis. Conclusion These results suggested that recombinant plasmid and fusion protein can be as potential candidates for vaccine or an antigen for diagnostic agents.
出处
《科技通报》
北大核心
2004年第2期172-174,177,共4页
Bulletin of Science and Technology
关键词
肺炎衣原体
膜表面蛋白
重组质粒
Chlamydia pneumoniae
outer membrane protein
recombinant plasmid
