Construction of Eukaryotic Expressing Plasmid of New Chicken Interleukin 15 and Its Expression in vitro

A new type gene of chicken interleukin 15(NChIL-15) was amplified by gene splicing overlap extension PCR(SOE PCR).NChIL-15 was cloned into cloning vector pMD18-T,and the recombinant cloning plasmid was sequenced and named pMD18T-NChIL-15.NChIL-15 gene was then sequenced and cloned into eukaryotic expressing vector pcDNA3.1(+).The recombinant expressing plasmid pcDNA3.1(+)-NChIL-15 was identified and then transfected into 293T cell in vitro with Ca3(PO4)2.The expression of NChIL-15 protein was successfully detected by indirect immunofluorescence assay(IFA) using mouse-anti ChIL-15 monoclonal antibody.The research paved a way for the further study of NChIL-15 as an adjuvant.