Construction of mammalian co-expression plasmid pIRES-CEA-IL-2 and its expression in vitro / 中国癌症杂志

ABSTRACT

Purpose:To construct the mammalian co expression plasmid pIRES CEA IM 2 and detect the expression of the plasmid in vitro. Methods:Using cloning technique,the cDNA fragments of CEA gene and mouse molecule IL 2 gene were constructed into the mammalian co expression plasmid vector pIRES.The inserted target genes in the mammalian co expression plasmid were verified by nucleotide sequencing.C 26 cell line was transfected with this mammalian co expression plasmid using lipofecarnin reagent. The expression of CEA and IL 2 molecules were detected by ELISA technique.Results:The mammalian co expression plasmid pIRES CEA IL 2 was obtained by cloning technique. The nucleotide sequences of CEA gene and molecule IL 2 gene in this mammalian co expression plasmid had high homology with CEA standard strain (99.8%) and mouse IL 2 (99.9%) respectively. After transfection with this mammalian co expression plasmid, the CEA and IL 2 molecules were expressed in C 26 cells. Conclusions:The constructed mammalian co expression plasmid pIRES CEA IL 2 can express CEA and IL 2 molecules in vitro at the same time.