克隆了人质子感知受体TDAG8基因,构建了TDAG8基因的表达载体并瞬时转染293T细胞,检测了TDAG8的过表达.从人胃癌组织cDNA中克隆了TDAG8基因,亚克隆到pEASY-T1载体,通过测序、酶切、连接到表达载体pEGFP-N3,通过酶切、测序鉴定获得了pEGFP-N3-TDAG8重组质粒；表达载体以lipofectamine2000介导转染了293T细胞.Real time PCR法检测了外源基因在293T细胞中的表达.结果表明：克隆到了TDAG8基因并获得了pEGFP-N3-TDAG8表达载体,在293T细胞中检测到了表达过的TDAG8基因% To detect the over-expression of TDAG8 in transiently transfected 293T cells, human TDAG8 gene was cloned and the TDAG8 expressing vector was constructed. TDAG8 CDS sequence was cloned from human gastric cancer tissues and sub-cloned into the pEASY-T1 vector. By sequencing, digestion and ligation, TDAG8 was directionally cloned into eukaryotic expression plasmid pEGFP-N3. The reconstructed plasmid was identified with enzyme digestion and sequencing;the plasmid was transfected into 293T cells with lipofectamine2000. Real-time PCR was performed to assess exogenous TDAG8 expression in 293T cells. Our results indicate that the expression vector containing human TDAG8 gene has been established and TDAG8 gene has also been over-expressed successfully in 293T cells.
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