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为探索建立持续稳定表达 h BD- 2的哺乳类动物型工程细胞的可能性 ,作者研究构建真核重组表达载体 p CMV.tag2 B/h BD- 2以进行 COS- 7细胞转染表达实验。将本室克隆获得的 h BD- 2全长 c DNA片段插入带有报告基因的真核表达质粒 p CMV.tag2 B构建出 h BD- 2的重组表达载体 p CMV.tag2 B/h BD- 2 ,并经 DNA测序证明h BD- 2 c DNA片段的插入方向和其全长 c DNA的碱基组成顺序均准确无误。通过脂质体转染法将 p CMV.tag2 B/h BD- 2导入 COS- 7细胞。RT- PCR法和免疫印迹法分别从 m RNA水平和蛋白质水平检测到被转染的 COS- 7细胞系能有效表达 h BD- 2。
To explore the possibility of establishing a mammalian engineering cell that can stably express h BD-2, we constructed an eukaryotic recombinant expression vector pCMV.tag2 B / h BD-2 for COS-7 cell transfection and expression assay. The full-length cDNA fragment of h BD-2 cloned in our laboratory was inserted into the eukaryotic expression plasmid pCMV.tag2 B with the reporter gene to construct a recombinant expression vector h BD-2 p CMV.tag2 B / h BD-2 , And confirmed by DNA sequencing h BD-2 c DNA insertion direction and its full-length c DNA base sequence composition are accurate. P CMV.tag2 B / h BD-2 was introduced into COS-7 cells by lipofection. RT-PCR and Western blotting confirmed that the transfected COS-7 cell line could express h BD-2 efficiently from the level of m RNA and the level of protein, respectively.