VR1012-TCRγV_1真核表达载体的构建

Construction of eukaryotic expression vector VR1012-TCRγV_1

目的:构建含TCRγV1基因的重组表达载体。方法:从人淋巴瘤Jurkat细胞中提取总RNA,反转录合成 cDNA克隆入VR1012质粒;转化感受态细菌E.ColiDH5α,以通用引物PCR扩增筛选阳性克隆,并双酶切鉴定;小 量提取质粒进行测序。结果:测序结果证实重组载体中的外源片段序列与GenBank中TCRγV1序列完全相符。结 论:成功构建了含TCRγV1基因的重组表达载体。

Aim: To construct the eukaryotic expression vector VR1012-TCRγV_1. Methods: The total RNA was extracted from human Jurkat cells;followed by a RT step to synthesize cDNA and a PCR amplification.The segment was cloned into VR1012. The recombinant was detected by PCR,endonuclease and sequencing. Results: The sequenced segment in the recombinant perfectly coincided with the selected sequence. Conclusion: The eukaryotic expression vector containing TCRγV_1 gene is constructed successfully.