沙眼衣原体主要外膜蛋白21-387的原核表达及其免疫原性

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Abstract Objective: To explore prokaryotic expression and immunogenicity of major outer membrane protein (MOMP21-387) of Chlamydia trachomatis serotype E. Methods: The gene encoding MOMP21-387 was amplified from genome DNA of C. trachomatis E by PCR analysis, and then cloned into pET21a (+) vector to construct recombinant plasmid pET21a (+)/MOMP21-387, and was expressed in the prokaryotic expression system. The MOMP21-387 fusion protein was purified and identified by SDS-PAGE and western blot analysis. The immunogenicity was further assessed by immunizing BABL/c mice, the reactivity of specific serum IgG in serum and genital tract mucosal IgA were tested by ELISA, and the specific cytotoxicity of spleen cells was detected with lactate dehydrogenase (LDH) method. Results: The MOMP21-387 fusion protein was successfully expressed in a prokaryotic expression system, and the specific positive bands at the relative molecular mass (Mr) of about 44 000 was confirmed by SDS-PAGE and western blot analysis; And the purified MOMP21-387 fusion protein was obtained with the Ni-NTA affinity chromatography method. The mice can be induced to produce the specific serum IgG and reproductive tract mucosal IgA by immunizing with the MOMP21-387 fusion protein, and the value of the specific serum IgG and mucosal IgAin immunized groups were significantly higher than that of the PBS control group (P<0.05), the antibodys detected by ELISA reached peak at the 6th week post-immunization. The LDH analysis showed that, the killing rate of spleen cells to the target cell in MOMP21-387 protein immunized groups was significantly higher than those in PBS control group, when the effector cells to target ratio reached to 10:1, 20:1, 40:1 and 80:1 (P<0.05). Conclusion: The MOMP21-387 fusion protein of Chlamydia trachomatis serovar E shows good immunogenicity, It may lay the foundation for the Chlamydia trachomatis ELISA detection and vaccine development based on MOMP21-387.

Key words： Chlamydia trachomatis major outer memberane protein prokaryotic expression immunogenicity

Received: 21 July 2014

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	LIU Qiaoqiong
	TU Jianxin
	LIN Xiaoyun
	XIONG Yirong
	ZHU Shanli
	CHEN Shao
	ZHANG Lifang.

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https://xb.wmu.edu.cn/EN/ OR https://xb.wmu.edu.cn/EN/Y2015/V45/I2/95

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