Expression of human B-7.1 transgene cell line and detection of the expressing pattern

Objective Construction of B-7.1 eukaryotic expression vector and import CAK-1 expression in renal cell carcinoma,for further study of gene function to do material preparation.Methods By RT-PCR method,regarding normal liver tissue cDNA as a template,B-7.1 gene is amplified full-length reading frame.And then construct the eukaryotic expression vector pCDNA3.1,import it to CAK-1 renal cell carcinoma cells,The transfected cells was identified by RT-PCR and Western blot analysis.Results the recombinant was verified by DNA sequencing,and the expression pattern of protein was identified by RT-PCR with the transcriptom of cells after transfection with pCDNA-B7 vector.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 Construction of human B-7.1 gene eukaryotic expression vector and detection of the expressing pattern provides ideal material in studying on the functional role of B-7.1 protein in tumorigenesis.