Construction of eukaryotic expression vector of human interleukin-24 and its expression in HepG2 cells

Abstract

Objective To construct a eukaryotic expression vector of human interleukin-24 (hIL-24) and to observe its expression in HepG2 cells. Methods The hIL-24 gene was cloned from phytohemaggluti-nin-treated human peripheral blood mononuclear cells with RT-PCR. The eukaryotic expression vector pcDNA3.1( + )-IL-24 was constructed by subcloning IL-24 cDNA into the eukaryotic expressing vector pcDNA3.1( + ) with DNA recombination technique and was identified by PCR, restriction enzyme digestion and DNA sequencing. The pcDNA3. 1 ( + )-IL-24 was stably transfected into HepG2 cells by liposome transfection with using G418 to screen positive transformants. RT-PCR was performed to determine hIL-24 mRNA expression in HepG2 cells. Results The target gene with expected molecular mass was obtained with RT-PCR. The results of PCR, restriction enzyme digestion and DNA sequencing showed that the IL-24 gene had cloned into the eukaryotic vector pcDNA3.1( + ) correctly. The mRNA of IL-24 was detected in the stably transfected HepG2 cells. Conclusions The recombinant eukaryotic expression vector pcDNA3.1 ( + )-IL-24 is constructed successfully and the HepG2 cell strains stably transfected with the recombinant plasmid are obtained. Key words: Interleukins; Gene expression; Vectors; HepG2 cells