摘要
Objective: To construct a recombinant plasmid containing the outer membrane protein 2 (Omp2) gene of Chlamydia trachomatis and express Omp2 in E.coli. Methods: The omp2 gene of C. trachomatis serovar D was cloned into pQE30 vector following PCR amplification from genomic DNA. E. coli M15 transformants were induced to express the fusion protein by IPTG and the product was identified by SDS-PAGE and Western blot. Results: Confirmed by enzyme cleavage analysis and DNA sequencing, a correct recombinant plasmid pQE30/omp2 was constructed. The fusion protein from the transformants was approximately 60 kDa in size in SDS-PAGE analysis, which could specially react with anti-6 X His mouse monoclonal IgG antibodies. Conclusion: We successfully expressed Omp2 in E. coli M15, providing an efficient and simple system for assaying the immunological properties of Omp2.
目的构建含沙眼衣原体外膜蛋白2(Omp2)抗原编码基因的表达质粒,在E.coli中表达 Omp2。方法通过PCR方法扩增D型沙眼衣原体omp2基因片段,克隆于表达载体pQE30,转化大肠杆菌M15感受态细胞,IPTG诱导表达,用SDS-PAGE及Western blot进行Omp2表达的鉴定。结果经酶切分析和DNA序列鉴定,构建了pQE30/omp2重组表达质粒;获得了相对分子质量(Mr)约为60 × 103的融合蛋白,能与抗-6聚组氨酸单克隆抗体发生特异性反应。结论利用原核表达载体,成功地表达了Omp2,为进一步开展其免疫学功能的研究奠定了基础。
基金
This work was supported in part by grants from the Department of Science and Technology of Hunan Province (No. 01SSY2008-6) the Department of Health of Hunan Province (No. B2003-078).
