新型鸡白细胞介素15真核表达质粒构建及其体外表达  

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

作　　者：马德星[1] 李广兴[1] 马春丽[2] 杨静红[1] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030 [2]东北农业大学食品学院,黑龙江哈尔滨150030

出　　处：《中国家禽》2011年第18期12-15,共4页China Poultry

基　　金：国家自然科学青年基金项目(30901061);东北农业大学科学研究基金(2010RCB38)

摘　　要：应用重叠扩增PCR(SOEPCR)技术将鸡白细胞介素15(ChIL-15)的冗长信号肽序列替换为较短的鸡白细胞介素2(ChIL-2)信号肽序列,构建了一种新型ChIL-15融合基因(NChIL-15)。将其连接到克隆载体pMD18-T中得到重组克隆质粒pMD18T-NChIL-15。阳性克隆质粒经鉴定正确后将NChIL-15基因亚克隆入真核表达载体pcDNA3.1(+)中,得到重组阳性表达质粒pcDNA3.1(+)-NChIL-15。阳性表达质粒经鉴定正确后应用磷酸钙法体外转染293T细胞,通过间接免疫荧光试验(IFA)检测到了NChIL-15蛋白在293T细胞中的表达。本研究为NChIL-15佐剂作用的深入研究奠定了基础。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.

关 键 词：NChIL-15 转染 间接免疫荧光试验 

分 类 号：Q782[生物学—分子生物学]