弓形虫GRA7真核表达质粒的构建

Construction of Eukaryotic Recombinant Expression Plasmid Congtaining Toxoplasma Gondii GRA7

目的:构建弓形虫致密颗粒抗原GRA7的真核重组表达质粒。方法:设计GRA7的特异引物,采用多聚酶链反应(PCR)技术从弓形虫RH株基因组DNA中扩增编码GRA7的基因片段,经克隆至pMD18-T载体后,亚克隆至真核表达载体pVAX1而构建真核重组表达质粒pVAX1-GRA7。结果:PCR扩增出GRA7基因的特异片段,所获克隆的序列正确,并被亚克隆到真核表达载体pVAX1,构建了真核重组表达质粒pVAX1-GRA8。结论:成功构建了GRA7的真核重组表达质粒pVAX1-GRA7。

Objective To construct an eukaryotic expression plasmid pVAX1-GRA7 containing the gene encoding Toxoplasma gondii RH strain dense granule antigen GRA7.Methods The gene fragment encoding GRA7 was amplified by PCR with special primers from Toxoplasma gondii genomic DNA,and was cloned into pMD18-T vector;The right gene fragment encoding GRA7 in positive clone was digested with Xba Ⅰand incompletively digested with Hind Ⅲ,and was subcloned into pVAX1 which was also digested with Xba Ⅰand Hind Ⅲ;the recombinant ...