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Position: Home > Articles > Construction and Identification of the Eukaryotic Expression Plasmid Containing GRA 3 Gene of Toxoplasma gondii Journal of Henan Agricultural Sciences 2010,39 (6) 132-135

弓形虫GRA3基因真核表达质粒的构建与鉴定

作  者:
薛书江;张守发;栾杨
单  位:
延边大学农学院
关键词:
弓形虫;GRA3基因;真核表达;鉴定
摘  要:
以pGEX-GRA3为模板,经PCR扩增获得弓形虫GRA3基因,克隆于pMD-18T simple载体上。重组质粒经PCR鉴定、PstⅠ和XhoⅠ双酶切鉴定后测序分析,再将其定向插入真核表达载体pVAXⅠ,构建真核表达重组质粒pVAX-GRA3。经PCR和酶切鉴定后,将重组质粒pVAX-GRA3转染Hela细胞,然后进行RT-PCR检测。RT-PCR结果表明,检测到了目的基因的存在,说明pVAX-GRA3成功转入Hela细胞。
译  名:
Construction and Identification of the Eukaryotic Expression Plasmid Containing GRA 3 Gene of Toxoplasma gondii
作  者:
XUE Shu-jiang,ZHANG Shou-fa,LUAN Yang(Agricultural College of Yanbian University,Longjing 133400,China)
关键词:
Toxoplasma gondii;GRA 3 gene;Eukaryotic expression;Identification
摘  要:
The GRA 3 gene of Toxoplasma gondii was amplified from plasmid pGEX-GRA3 by PCR and ligated into pMD-18T simple vector for DNA sequencing.Then,the PCR products were inserted into pVAXⅠvector to construct a recombinant plasmid named as pVAX-GRA3 and characterized by PCR and restriction-endonuclease PstⅠand XhoⅠdigestion.Hela cells were transfected by plasmid pVAX-GRA3 with lipofection strategy and the expression of GRA 3 gene were detected by RT-PCR.The results showed that we have successfully constructed a recombinant plasmid of GRA 3 gene of Toxoplasma gondii,which established a base for further study of vaccines using the GRA 3 gene.

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