关键词: 衣原体, 沙眼, 表位, T淋巴细胞, 聚合酶链反应, 方法, 重组融合蛋白质类

Abstract: Objective To construct the prokaryotic expression vector of HSP65-T cell epitopes of MOMP of C.trachomatis（Ct）fusion protein(called H-ctm1), express and purify H-ctm1. Methods The HSP65 and gene sequence of T cell epitopes of MOMP of Ct (ctm1) were obtained by PCR method and cloned into pMD18-T vector respectively. The T vector was digested by endonuclease for releasing the fragment of H SP65 or ctm1 gene. The gene of HSP65 and ctm1 were cloned into pET28a plasmid su ccessively to construct the prokaryotic expression vector (pET28a-H-ctm1), whi ch recombined the gene of HSP65 and ctm1. The E.coli BL21(DE3) transformed with pET28a-H-ctm1 were induced by IPTE for expression of H-ctm1 fusion protein. The protein was purified by Ni²⁺ affinity chromatography. Results The prokaryotic expression vector, pET28a-H-ctm1, was constructed successfully. The fusion protein of H-ctm1 was expressed in E.coli BL21(DE3). The purity of fusion protein was 98% after purified by Ni²⁺ affinity chromatography. Conclusion The prokaryotic expression vector of pET28a-H-ctm1 has been constructed, and the fusion protein with biological activity has been successfully expressed and purified.

Key words: Chlamydia trachomatis, epitopes, T-lymphocyte, polymerase chain reaction, methods, recombinant fusion proteins