Expression and Identification of Major Outer Membrane Protein of Chla mydia trachomatis in Escherichia coli
- VernacularTitle:沙眼衣原体主要外膜蛋白在大肠杆菌中的表达与鉴定
- Author:
Zhongyu LI
;
Yimou WU
;
Chaoqun CHEN
;
Minjun YU
- Publication Type:Journal Article
- Keywords:
Chlamydia trachomatis;
Bacterial outer membrane proteins;
Gene expression
- From:
Chinese Journal of Dermatology
2003;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the recombinant plasmid containing the major outer membrane protein(MOMP) gene of Chlamydia trachomatis and expres s MOMP protein in E.coli BL21. Methods The MOMP gene was amplified by polymera se chain reaction from the genome of Chlamydia trachomatis serovar D. The amplif ied fragment was directly inserted into pUCm-T vector and verified by DNA sequen cing. MOMP gene was then subcloned into the prokaryotic expression vector pET-22 b(+). The recombinant protein of MOMP was purified by Ni-NTA affinity chromatogr aphy and identified by SDS-PAGE and Western blot. Results The MOMP gene, which is about 1 200 bp, was successfully amplified and cloned. The DNA sequence of t he cloned MOMP gene was the same as that published by the GenBank. SDS-PAGE anal ysis showed that the relative molecular weight of this fusion protein was about 47 kDa which was consistent with the theoretically predicted value, and the spec ificity of this recombinant protein was confirmed by Western blot. Conclusions The MOMP gene of Chlamydia trachomatis was successfully cloned and expressed in the prokaryotic expression system, which may lay the foundation for the developm ent of Chlamydia trachomatis vaccine.
