Construction of the plasmid expressing gfp and MAD2 gene controlling separation of cell and its expression in HepG2 cell / 重庆医科大学学报

ABSTRACT

Objective: To establish an in vitro screening system for testing effects of MAD2 gene of embryonic cell genome.Methods: In order to construct the plasmid expressing MAD2-GFP fusion plasmid,the target gene fragment was obtained by RT-PCR amplification and inserted into pEGFP-N1 reporter vector.The structure of constructed plasmid was confirmed by electrophoresis analysis and DNA sequencing.The function of construct was confirmed by lipofectamine-mediated transient expression in HepG2 cells.Results: DNA sequencing showed that the inserted fragment of the constructed plasmid was the same as the template MAD2 genome.The HepG2 cells transfected with the constructed plasmid could express reporter gene of gfp.Conclusions: the plasmid expressing gfp gene controlled by MAD2 gene is successfully constructed and an in vitro testing system for evaluating founction of MAD2 gene in separation of cell is established.