Construction and expression of eukaryotic plasmids containing human IL-27 protein subunits

Objective To clone and construct the eukaryotic expression plasmids containing human IL-27 protein subunits and investigate their expression following transfection into COS-7 cells. Methods Total RNA was extracted from dendritic cells derived from human peripheral blood monocytes for cloning EBI3 cDNA and p28 cDNA by RT-PCR.The eukaryotic expression plasmids pcDNA-EBI3 and pcDNA-p28 were constructed and transfected into COS-7 cells,where the expressions of target gene were detected by RT-PCR.Results The IL-27 protein subunits cDNA were cloned and the constructed plasmids were verified to be correct by sequencing.Target gene expressions were detected by RT-PCR in the transfected COS7 cells.Conclusion The eukaryotic expression plasmids containing human IL-27 protein subunits were constructed and expressed in COS-7 cells successfully.This work paves the way for construction and expression of the bioactive single chain human IL-27 protein.