Construct the eukaryocytic expression vector of Id1 and transfection into immortal gastric mucosa epithelial cell

    HAN Shuang, DING Jie, GUO Changª²Cun, HAN Zheª²Yi, SHEN Huiª²Qin, CHEN Yu, ZHA Xicuomu, QIAO Taiª²Dong, WU Kaiª²Chun, FAN Daiª²Ming

    PLA Institute of Digestive Disease, Xijing Hospital, Fourth Military Medical University, Xi¬ðan 710033, China

    ¡¾Abstract¡¿ AIM: To amplify the Id1 gene and to construct a eukaryocytic expression vector of Id1. METHODS: RTª²PCR was performed to amplify the Id1 gene containing the complete CDS by using the total cDNA of gastric cancer cell line SGC7901 as template. Id1 gene was inserted into the pUCmª²T vector and sequenced. The correct Id1 cDNA was subcloned into eukaryocytic expression vector pcDNA3.1/V5HisA and transfected into GESª²1 by lipofectamineª²mediated transfection. RESULTS: RTª²PCR yielded a fragment of 608 bp and Id1 was verified by sequence analysis. The subclone was identified by restriction enzyme digestion. Western blotting confirmed the transfetant in which Id1 was over expressed. CONCLUSION: Eukaryocytic expression vector of Id1 has been successfully constructed, which provides a premise for further investigation of its function in the gastric cancer. ......