Cloning and sequence analysis genes of porcine interleukin-12 p35 and p40 subunit

Objective: To clone genes of encoding porcine interleukin-12 p35 and p40 subunits,for preparing the compound nucleir acid vaccine against the porcine bladder worm.Methods: Peripheral blood mononuclear cells and splenic lymphocytes of porcine were isolated and cultured for 10 hours,and stimulated with 100 ng/ml IFN-γ for 12 hours,followed by stimulated with 1 μg/ml lipopolysaccharide for 3 hours.Total RNA from these stimulated porcine lymphocytes were extracted,cDNA of genes for porcine IL-12 p35 and p40 subunit were ampliphied by RT-PCR assay and inserted into pMD18-T vector.The cloned genes were sequenced.Results: The PCR products of porcine IL-12 p35 and p40 subunits were 616 bp and 1 011 bp,respectively,verified by electrophoresis,sequence analysis for cloned genes in pMD18-T vector showed one base was different in each but none affecting amino acids coding,confirmed to be genes of porcine IL-12 p35 and p40 subunit.Conclusions: cDNA encoding porcine interleukin-12 p35 and p40 subunit have been successfully cloned into cloning vector pMD18-T.