Cloning of human CD81 and its expression in COS-7 cells

Aim To clone human CD81 gene from peripheral blood lymphocytes, and construct its eukaryotic expression vector, and then express it in COS-7 cells. MethodsCD81 gene was amplified by RT-PCR,using RNA extracted from peripheral blood lymphocytes as template, and was cloned into pcDNA3.1(+) vector .Constructed pcDNA3.1-CD81 was identified by endonucleases digestion and sequencing. The recombinant expression plasmid was transfected into COS-7 cells. Human CD81 expressed on membrane of COS-7 cells was detected by immunocytochemical staining and FACS. ResultsObtained full encoding sequence of CD81 was identitical with that included in Genbank, and the eukaryotic expression vector pcDNA3.1-CD81 was constructed correctly. Expression of human CD81 on COS-7 cells was proved by the immunological detection. ConclusionThe eukaryotic expression plasmid pcDNA3.1-CD81 was constructed successfully, which will contribute to further studies on interaction of CD81 with HCV and roles of CD81 in the process of HCV infection and to the establishment of possible better HCV cellular infection model.