山羊TLR2绿色荧光蛋白融合载体的构建

Construction of TLR2 fusion vector of green fluorescent protein in goat

为了构建在哺乳动物细胞中表达的山羊TLR2绿色荧光蛋白融合载体,试验根据克隆到T载体上的山羊TLR2测序结果,设计1对不含终止子的引物,将山羊TLR2 PCR产物连接到pEGFP-N1载体上,用磷酸钙法转染293T细胞,并在荧光显微镜下观察。结果表明:重组质粒经酶切和测序鉴定正确,且在293T细胞中表达;融合蛋白发出绿色荧光,表明TLR2-EGFP主要分布在细胞膜上。说明试验成功地构建了pEGFP-TLR2-N1绿色荧光蛋白融合载体。

To construct the fusion expression vector of goat TLR2 fused to green fluorescent protein which is capable of expression in mammalian cells.A pair of primers was designed based on the sequence of T-TLR2 vector.The goat TLR2 was cloned into the green fluorescent protein vector pEGFP-N1.The recombinant vector was then transfected into 293T cells,and the cells were observed by using fluorescent microscope.The recombinant vector was confirmed to be constructed successfully identified by enzyme digestion and sequencing,which was highly expressed in 293T cells.The fusion protein of TLR2-EGFP was observed mainly in the cell membrane.The expression vector of pEGFP-TLR2-N1 fused to green fluorescent protein is successfully constructed,which may facilitate to further study the application of TLR2.