MYCT-1基因真核表达载体的构建及鉴定

目的构建MYCT-1(myc target)基因真核表达载体,观察其转染胃粘膜GES-1细胞系后的表达。方法用RT-PCR法合成MYCT-1 cDNA,克隆至表达载体pcDNA3.1上,测序验证无突变发生。将表达载体转染GES-1细胞,用RT-PCR和Western blot检测MYCT-1基因的表达。结果限制性内切酶酶切和DNA测序证实成功克隆了MYCT-1 cD-NA全长并正确插入了表达载体,在GES-1细胞中检测到MYCT-1 mRNA和蛋白。结论成功构建了MYCT-1真核表达载体,可在GES-1细胞中稳定表达。

Construction and identification of an eukariotic expression vector of MYCT-1

Objective To construct an eukariotic expression vector of MYCT-1 and detect its expression after transfected in GES-1 cells.Methods MYCT-1 cDNA was synthetized by RT-PCR and then cloned into an expression vector pcDNA3.1.DNA sequencing was performed to avoid any mutation in recombianant vector.The vector pcDNA3.1-MYCT-1 was transfected into GES cells by liposome and the expression of MYCT-1 was detected by RT-PCR and Western blot.Results The full length cDNA of MYCT-1 cloned into the exoression vector was i...