Construction of Human B-7.1 Gene Eukaryotic Expression Vector and Detection of the Expressing Pattern

Objective To subclone the human B-7.1 gene open reading frame(ORF)into eukaryotic expression vector pcDNA3.1 for investigating the biological function of B-7.1 protein.Methods RT-PCR was used to acquire the B-7.1 gene from normal liver tissue.PCR was done to amplify the ORF of B-7.1 and the PCR product Was digested and subcloned into pcDNA3.1 vector which had been digested by same restriction endonuclease.Results the recombinant was verified by DNA sequencing,and the expression pattern of protein was identified by RT-PCR with the transcription of cells after transfection with pcDNA-B7 vector.Conclusion Eukaryotic expression vector of B-7.1 gene provides ideal material in studying on the functional role of B-7.1 protein in tumor genesis. Key words: B-7.1;  Vector;  Eukaryotic expression