Cloning, expression and phylogenetic analysis of IFN- gamma gene in chicken.

摘要

In the present study, chicken interferon gamma (Ch.IFN- gamma ) gene was amplified using specific primers by RT-PCR and cloned into pTZ 57R/T cloning vector. The clone was confirmed by restriction digestion analysis using KpnI and ApaI restriction enzymes and orientation was confirmed by colony PCR using T7 promoter primer and gene specific forward primers. Further, the gene fragment was sub-cloned in eukaryotic expression vector pcDNA 3.1(+) and confirmed by restriction digestion. The recombinant pcDNA 3.1 having IFN gene (pcDNA.Ch-IFN- gamma ) was checked for its ability to express IFN- gamma in vitro in cell culture and expression was confirmed by RT-PCR, indicating the gene is functionally active. The study to evaluate ability of pcDNA.Ch-IFN- gamma to combat infection and enhance efficacy of DNA vaccine is under way.