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目的:构建TRBP及其截短体真核表达载体,并检测其在HeLa细胞中的表达。方法:设计引物扩增TRBP全长及其截短体TAB、TAC、TBC基因序列,然后将各基因片段克隆入真核表达载体pFlag-CMV4。用脂质体法转染HeLa细胞,经Western blot方法检测目的基因在转染细胞中的表达。结果:经过酶切鉴定与测序证实,TRBP全长及其截短体TAB、TBC、TAC基因真核表达载体构建成功,经Western blot可检测转染细胞中各基因的表达。结论:成功构建了TRBP全长基因及其截短体TAB、TAC和TBC基因真核表达载体,在转染细胞中可以检测到目的基因的表达。
OBJECTIVE: To construct the eukaryotic expression vector of TRBP and its truncated trx, and to detect its expression in HeLa cells. Methods: Primers were designed to amplify the full-length TRBP and its truncated TAB, TAC and TBC sequences, and then cloned into the eukaryotic expression vector pFlag-CMV4. HeLa cells were transfected by liposome method and the expression of target gene in transfected cells was detected by Western blot. Results: The eukaryotic expression vectors of TAB, TBC and TAC were successfully constructed by restriction endonuclease digestion and sequencing. The expression of each gene in transfected cells was detected by Western blot. CONCLUSION: The full-length TRBP gene and its truncated TAB, TAC and TBC gene eukaryotic expression vector were successfully constructed, and the expression of the target gene can be detected in transfected cells.