Construction of the eukaryotic expression vector containing the human perforin gene and Its expression in COS-7 cells / 医学研究生学报

ABSTRACT

Objective: To clone human perforin(pfn) full-length DNA,construct the eukaryotic expression vector and observe the expression of the pfn gene in the transfected COS-7 cells.Methods: Full-length DNA of pfn was obtained by PCR from rPCR2.1/pfn,inserted into the pMD-18T vector and subcloned to the pcDNA3.1(+) vector to construct the recombinant eukaryotic expression vector pcDNA3.1(+)/pfn.The recombinant plasmid was transfected into COS-7 cells and the expression of the pfn gene in the transfected cells was detected by RT-PCR.Results: The pfn full-length DNA was successfully cloned and inserted into the pcDNA3.1(+) vector.The expression of pfn mRNA in the transfected COS-7 cells was confirmed by RT-PCR. Conclusion: The full-length human pfn gene can be expressed in transfected COS-7 cells.