山羊BST-2基因的表达及其亚细胞定位  

Expression and subcellular localization of bone marrow stromal antigen 2 gene of Capra hircus

作　　者：张莉[1,2] 刘腾 缪秋红[2] 唐井玉[2] 朱杰 董丹丹[2] 陈宗艳 王桂军[1] 刘光清[2] ZHANG Li1,2,LIU Teng 2,MIAO Qiuhong2, TANG Jingyu2,ZHU Jie2,DONG Dandan2,CHEN Zongyan2, WANG Guijim1 ,LIU Guangqing2(1. College of Animal Science and Technology,Anhui Agricultural University,Hefei 230036, China; 2. Shanghai Vet-erinary Research Institute , CAAS,Shanghai 200241,Chin)

机构地区：[1]安徽农业大学动物科技学院,安徽合肥230036 [2]中国农业科学院上海兽医研究所,上海200241

出　　处：《浙江农业学报》2018年第5期702-706,共5页Acta Agriculturae Zhejiangensis

基　　金：国家重点研发计划(2016YFD0500108); 国家自然科学基金(31502068); 上海市科技兴农重点攻关项目(2016043); 公益性农业科研专项(201303046); 中央级公益性科研院所基本科研业务费专项(2016JB01)

摘　　要：将山羊的BST-2基因(Capra hircus bone marrow stromal antigen 2 gene)分别克隆至原核表达载体p GEX-4T-1和真核表达载体p3×Flag-CMV-14中,获得了重组表达质粒p GEX-BST-2和p3×Flag-BST-2。将重组质粒p GEX-BST-2转化至E.coli BL21(DE3)感受态细胞中,用IPTG进行诱导表达。SDS-PAGE及Western blot结果显示,诱导表达的重组蛋白分子量约为42 ku,与预期蛋白一致。利用脂质体介导转染法将重组质粒p3×Flag-BST-2转染Vero细胞,经激光共聚焦显微镜检测,结果显示,BST-2蛋白在细胞中可以良好表达,并主要定位于细胞膜上。BST-2 gene（ Capra hircus bone marrow stromal antigen 2 gene） was amplified and cloned into p GEX-4 T-1 and p3 × Flag-CMV-14,respectively. The recombinant plasmids were named as p GEX-BST-2 and p3 × Flag-BST-2,respectively. Firstly,p GEX-BST-2 was transformed into E. coli BL21（ DE3）,then the protein expression was induced by IPTG. The results of SDS-PAGE and Western blot analysis showed that the molecular weight of the recombinant protein was about 42 ku. Finally,Vero cells were transfected with p3 × Flag-BST-2 through Lipofectamine2 000 reagent,and the cells were analyzed by confocal microscopy.

关 键 词：BST-2基因 原核表达 真核表达 亚细胞定位 山羊 

分 类 号：S827[农业科学—畜牧学]