Expression and Detection of Human B7-1 Transgene Cell Line

Objective To construct B7-1 eukaryotic expression vector and import CAK-1 expression in renal carcinoma cells and to prepare materials for further study of gene function.Methods By reverse transcription polymerase chain reaction(RT-PCR) method,normal liver tissue cDNA was utilized as template,B7-1 gene full-length reading frame was amplified,the eukaryotic expression vector pCDNA3.1 was constructed and imported to CAK-1 renal carcinoma cells.The transfected cells were identified by RT-PCR and Western blot analysis.Results The cloning and sequencing results demonstrated that B7-1 reading frame was successfully inserted the corresponding site of eukaryotic expression vector pCDNA3.1.Compared with pCDNA3 empty vector control cells,the B-7 mRNA and protein levels were significantly higher in the pCDNA3-B-7 vector-transfected cells.Conclusion Successful construction and detection of human B7-1 gene eukaryotic expression vector provides the ideal material for further study of the functional role of B7-1 protein in tumorigenesis.