| 山羊BST-2基因的表达及其亚细胞定位 |
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| 引用本文： | 张莉,刘腾,缪秋红,唐井玉,朱杰,董丹丹,陈宗艳,王桂军,刘光清.山羊BST-2基因的表达及其亚细胞定位[J].浙江农业学报,2018,30(5):702. |
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| 作者姓名： | 张莉 刘腾 缪秋红 唐井玉 朱杰 董丹丹 陈宗艳 王桂军 刘光清 |
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| 作者单位： | 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 |
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| 基金项目： | 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) |
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| 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pGEX-4T-1和真核表达载体p3×Flag-CMV-14中,获得了重组表达质粒pGEX-BST-2和p3×Flag-BST-2。将重组质粒pGEX-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。
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| 关 键 词： | BST-2基因 原核表达 真核表达 亚细胞定位 山羊 |
| 收稿时间： | 2017-07-12 |
Expression and subcellular localization of bone marrow stromal antigen 2 gene of Capra hircus |
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| ZHANG Li,LIU Teng,MIAO Qiuhong,TANG Jingyu,ZHU Jie,DONG Dandan,CHEN Zongyan,WANG Guijun,LIU Guangqing.Expression and subcellular localization of bone marrow stromal antigen 2 gene of Capra hircus[J].Acta Agriculturae Zhejiangensis,2018,30(5):702. |
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| Authors: | ZHANG Li LIU Teng MIAO Qiuhong TANG Jingyu ZHU Jie DONG Dandan CHEN Zongyan WANG Guijun LIU Guangqing |
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| Institution: | 1.College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China;
2.Shanghai Veterinary Research Institute, CAAS, Shanghai 200241, China |
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| Abstract: | BST-2 gene (Capra hircus bone marrow stromal antigen 2 gene) was amplified and cloned into pGEX-4T-1 and p3×Flag-CMV-14, respectively. The recombinant plasmids were named as pGEX-BST-2 and p3×Flag-BST-2, respectively. Firstly, pGEX-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 Lipofectamine 2 000 reagent, and the cells were analyzed by confocal microscopy. |
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| Keywords: | BST-2 gene (bone marrow stromal antigen 2 gene) prokaryotic expression eukaryotic expression subcellular localization Capra hiracs |
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