摘要
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CtMOMP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。结果对构建的重组菌以种特异性抗CtMOMP单克隆抗体(mAb)做蛋白印迹表明 ,在相对分子质量(Mr)约为33000处表达有1条带 ,表达量约占菌体总蛋白的13 %。结论成功地构建了能表达MOMP的重组菌株并获得表达。
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.
出处
《细胞与分子免疫学杂志》
CSCD
2000年第4期311-313,共3页
Chinese Journal of Cellular and Molecular Immunology
关键词
沙眼衣原体
MOMP
减毒鼠伤寒杆菌
基因表达
Chlamydia trachomatis
major outer membrane pro tein
attenuated Salmonella typhimurium
gene c
loning
gene expression
