摘要
目的对F型沙眼衣原体主外膜蛋白(MOMP)基因进行原核表达,获得omp1基因重组蛋白,并对克隆到的基因进行序列分析,为F型沙眼衣原体的诊断及疫苗研究奠定基础。方法利用PCR技术从沙眼衣原体阳性病人尿道拭子标本中扩增omp1基因,分析序列,并定向克隆入原核表达载体pGEX-5X3中,与GST进行融合表达。结果对克隆到的沙眼衣原体omp1基因进行序列分析,基因序列与F型沙眼衣原体F/IC-CAL3相似性达99%,构建Omp1-pGEX-5X3重组质粒,omp1与GST融合表达。结论成功构建F型沙眼衣原体omp1基因原核表达系统,得到重组蛋白,为沙眼衣原体疫苗的研究和临床检测试剂盒的研制打下基础。
To express the gene encoding the major outer membrane protein （MOMP） of Chlamydia trachomatis serovar. F to obtain the ompl gene recombinant protein for use in diagnosis and vaccine development for C. trachomatis infection, the ompl gene was amplified from the genome of C. trachomatis from urethra swab sample of trachoma patients by PCR, sequenced and cloned to Prokaryotic expression vector pGEX-SX3 for fusion expression with GST. The cloned ompl gene was then subjected to sequence analysis and it was found that the sequence identity of ornpl gene with F/IC-CAL3 of C. trachornatis serovar F approached up to 99 %. Also, the Ompl-pGEX-SX3 recombinant plasmid was successfully constructed. It is apparent that the prokaryotic expression system for the ompl gene is established and the recombinant protein obtained, thus providing the foundation for the studies on trachoma vaccine and preparation of the diagnostic kids for trachoma.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2008年第11期1052-1056,共5页
Chinese Journal of Zoonoses
基金
国家自然科学基金(No.30400258)
关键词
F型沙眼衣原体
omp1基因
原核表达
序列分析
Chlamydia trachomatis serovar F
ompl gene
prokaryotic expression
sequence analysis
