刚地弓形虫Rop18基因的原核表达及鉴定

Abstract

Abstract: In this study,the Rop18 gene of T.gondii was cloned and expressed in E.coli Top10, and the immunogenicity of the recombinant protein was analyzed. The coding region of Rop18 was amplified with PCR and cloned into the prokaryotic expression vector pET-30a. The recombinant plasmid were confirmed by PCR, enzyme digestion and DNA sequencing, which was then transformed into E.coli BL21 induced by IPTG.The expressed proteins were analyzed by SDS-PAGE and Western blotting.Results indicated that the recombinant pET30a-Rop18 was correctly constructed.The results of SDS-PAGE and Western blotting revealed that the molecular weight of the recombinant protein was approximately 66.2 ku and could be recognized by rabbit antiserum against T.gondii.The results suggested that the Rop18 gene of T.gondii could be expressed in E.coli,and immunogenicity of the recombinant protein had been identifieded by Western blotting indicating that it might be used as vaccine candidate antigen in the future.

Key words: Toxoplasma gondii; Rop18 gene; cloning; prokaryotic expression

CLC Number:

Q786

QU Dao-feng, HAN Jian-zhong, DU Ai-fang. Prokaryotic Expression of Toxoplasma gondii Rop18 Gene and Immunogenicity Analysis of the Recombinant Protein[J]. , 2013, 40(10): 72-75.

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Prokaryotic Expression of Toxoplasma gondii Rop18 Gene and Immunogenicity Analysis of the Recombinant Protein

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