弓形虫表面抗原P22基因片段的克隆、表达及鉴定

CLONE,EXPRESSION AND IDENTIFICATION OF THE RECOMBITANT SURFACE ANTIGEN P22 GENE FRAGMENT OF TOXOPLASMA GONDII

目的 扩增弓形虫表面抗原P2 2基因编码序列 ,并进行表达和鉴定。 方法 设计合成引物 ,PCR法从RH株弓形虫基因组DNA中扩增P2 2基因编码序列 ,克隆入载体pET 3 2a ,转化大肠埃希菌BL2 1,IPTG诱导表达 ,表达产物进行SDS PAGE和Westernblot鉴定。 结果 从弓形虫基因组DNA中扩增出P2 2基因编码序列 ,并诱导表达出能被兔抗弓形虫血清识别的重组P2 2。 结论 成功获得弓形虫表面抗原P2 2的表达产物 ,为弓形虫病的诊断和疫苗研究创造了条件。

Objective To clone,express and identify major surface antigen P22 gene fragment of Toxoplasma gondii . Methods The P22 gene fragment was amplified from genomic DNA of T. gondii RH strain by PCR. The amplified fragment was cloned into the vector pET-32a,converted into Escherichia coli BL21,then expression was induced by IPTG. The expression product was analyzed by SDS-PAGE and western blot. Results The P22 gene fragment of T. gondii was amplified .The result of western blot analysis showed that the recombinant P22 can be recognized by the serum of rabbit immunized with T. gondii antigen. Conclusion The P22 fusion protein was successfully obtained and it might be used in immunological diagnosis and vaccination of toxoplasmosis.