Construction of co-expression vector of pVAX1-PIG and expression of granzyme B and perforin in human Hep-2 cell line / 中国免疫学杂志

ABSTRACT

Objective:To amplify full-length expressing sequence of human granzyme B(GrB) and perforin(PFP) firstly,then to construct a co-expression vector of pVAX1-PIG and to analyze the expression of human GrB and PFP in Hep-2 cells.Methods:The human PFP and GrB expressing sequences were amplified by RT-PCR.The expression vector pVAX1-PIG was constructed.The recombinant vector was transfected into human Hep-2 cells and their expressions were detected by RT-PCR and indirect immunofluorescent antibody assay.Results:The GrB and PFP cDNA fragment were cloned into the vector of pVAX1 in the right direction.The target proteins were detected in the transfected Hep-2 cells.Conclusion:The vector of pVAX1-PIG was constructed successfully and expressed in Hep-2 cells line.The perforin/granzyme B protein can induce cell apoptosis.These results provide some foundation in gene therapy for tumor by making use of PFP and GrB gene.