安徽医科大学学报2001，Vol.36Issue(1)：12-14,3.
弓形虫14-3-3信号转导蛋白基因亚克隆及鉴定
Subcloning and identification of 14-3-3 signal transduction protein gene of toxoplasma gondii
摘要
Abstract
Objective 14-3-3 signal transduction protein gene of Toxoplasma gondii(Toxo14-3-3) was subcloned into eukaryotic expression vector pBK-CMV for characterization of a new candidate vaccine and diagnostic molecules and for studies of the interaction between parasite and host cells. Methods A pair of primers were designed and synthesized based on the DNA sequence cloned from the enteroepithelial stage of Toxoplasma gondii. The DNA fragment encoding Toxo14-3-3 was amplified by RT-PCR from RNA of RH strain tachyzoites. The purified PCR products were ligated to T-vector pGEM-T. The inserted DNA fragment was confirmed by DNA sequencing. Then the target gene was subcloned into eukaryotic expression vector pBK-CMV. E.coli XL1-blue strain was transformed and the transformants were screened with LB Kanamycin media. The recombinant vector pBK-CMV /Toxo14-3-3 was identified by restriction endonuclease digestion and PCR. Results A open reading frame(ORF) of 798 bp was verified,which codes 265 amino acids. Conclusion A positive recombinant clone pBK-CMV /Toxo14-3-3 was obtained which lays a foundation for further expression recombimant.关键词
弓形虫属/基因表达/克隆,分子/信号转递分类
医药卫生引用本文复制引用
王维,汪学龙,沈继龙,蒋作君..弓形虫14-3-3信号转导蛋白基因亚克隆及鉴定[J].安徽医科大学学报,2001,36(1):12-14,3.基金项目
安徽医科大学青年科研基金资助项目 （）
