山羊天然抵抗力相关巨噬细胞蛋白基因(Nramp 1)的克隆及其在羊胎儿成纤维细胞中的表达和鉴定  

Expression of goat natural resistance associated macrophage protein gene in mice and sheep fetal fibroblast cells

作　　者：秦莹[1] 安星兰[1] 张学明[1] 岳占碰[1] 李子义[1] 

机构地区：[1]吉林大学动物医学学院动物胚胎工程吉林省重点实验室,吉林长春130062

出　　处：《中国兽医学报》2013年第3期481-485,共5页Chinese Journal of Veterinary Science

基　　金：国家转基因生物新品种培育科技重大专项(2011ZX08008-005);国务院华侨办华人创业团队基金重点专项资助项目

摘　　要：Nramp1(Slc11a1)表达于晚期吞噬溶酶体中,能够抵抗多种细胞内病原体的感染。Nramp1主要是通过调节巨噬细胞的免疫功能和细胞铁代谢发挥抗菌作用。为了研究山羊天然抵抗力相关巨噬细胞蛋白的功能,寻找一种在山羊中抵抗胞内菌感染的新途径。通过RT-PCR的方法从山羊脾脏中克隆Nramp1基因,连接到真核表达载体pcDNA3.1(+)上,命名为pcDNA3.1(+)-Nramp1。取60d小尾寒羊胎儿,建立胎儿成纤维细胞系,将线性化的载体转染胎儿成纤维细胞,获得稳定转染的细胞系,在基因组和转录水平分别对转基因细胞系进行鉴定,结果显示外源基因成功整合到基因组中并且在转录水平检测到基因表达,最后获得阳性转基因细胞系2株,为Nramp1蛋白的功能研究和抗菌试验奠定了基础。The expression of Nramp 1 （Slcl la 1） fection with several intracellular pathogens. The in late phagolysosomes confers resistance to in antimicrobial actions of Nramp 1 are attributa ble,in part, to modulation of macrophage immune function and cellular iron metabolism. In order to investigate the function of goat natural resistance associated macrophage protein and to find a effectively way on natural resistance to infection in goat. Nramp 1 gene was cloned from goat spleen by RT-PCR and was inserted into pcDNA3.1 （＋）, which named pcDNA3.1 （ ＋）-Nramp 1. Getting 60 days small tail han sheep fetal to establish fetal fibroblast cell lines while the linear- ized vector was transfected into sheep fetal fibroblast cells to gain stably transfected cell lines. However,the identification at the genome and transcription level shows that the exogenous gene successfully integrated into the genome and the gene expression is detected at transcription level. Two transgenic cell lines were carried out and laid a foundation of Nramp 1 protein function re- searching and antibacterial experiments.

关 键 词：山羊 NRAMP 1基因 真核表达 原核注射 

分 类 号：S852.2[农业科学—基础兽医学]