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 Chicken Interleukin 15 and Its Expression in vitro

作　　者：马德星[1] 潘龙[1] 杨静红[1] 盖仁华[1] 李广兴[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

出　　处：《中国家禽》2009年第6期8-11,15,共5页China Poultry

摘　　要：应用反转录-聚合酶链式反应(RT-PCR)技术从ConA刺激20h的白莱航鸡脾脏T淋巴细胞中扩增出鸡白细胞介素15(ChIL-15)全基因片段。将其连接到克隆载体pMD18-T中得到重组克隆质粒pMD18-T-ChIL-15。阳性克隆质粒经鉴定正确后将ChIL-15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(+)-ChIL-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ChIL-15蛋白在293T细胞中的表达。The complete gene fragment of Chicken interleukin 15 （ChIL-15）was amplified from T lymphocytes in spleen stimulated by ConA for 20 h by RT-PCR technique. Connect ChIL-15 to cloning vector pMD18-T,and the recombinant cloning plasmids were sequenced and named pMD18T-ChIL-15. ChIL-15 gene was then sequenced and cloned into eukaryotic expressing vector peDNA3.1 （＋）. The recombinant expressing plasmid was identified and transfeeted into 293T cell in vitro with Ca3 （PO4）2. 30 h after transfection,the expression of ChIL-15 protein was successfully detected by indirect immunofluoreseence assay （IFA） with mouse-anti ChIL-15 monoclonal antibody. The study paves the way for the further study of ChIL-15 as an adjuvant.

关 键 词：ChIL-15 转染 IFA 

分 类 号：Q782[生物学—分子生物学] Q78