山羊BST-2基因的表达及其亚细胞定位

doi:10.3969/j.issn.1004-1524.2018.05.04

浙江农业学报 ›› 2018, Vol. 30 ›› Issue (5): 702-706.DOI: 10.3969/j.issn.1004-1524.2018.05.04

山羊BST-2基因的表达及其亚细胞定位

张莉^1,2, 刘腾², 缪秋红², 唐井玉², 朱杰², 董丹丹², 陈宗艳², 王桂军^1,*, 刘光清^2,*

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

收稿日期:2017-07-12 出版日期:2018-05-20 发布日期:2018-05-23
通讯作者: 刘光清,E-mail: liugq@shvri.ac.cn ;王桂军,E-mail: wgj@ahau.edu.cn
作者简介:张莉(1992—),女,江苏苏州人,硕士,研究方向为动物传染病学。E-mail: 2385228064@qq.com
基金资助:
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)

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

ZHANG Li^1,2, LIU Teng², MIAO Qiuhong², TANG Jingyu², ZHU Jie², DONG Dandan², CHEN Zongyan², WANG Guijun^1,*, LIU Guangqing^2,*

1.College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China;
2.Shanghai Veterinary Research Institute, CAAS, Shanghai 200241, China

Received:2017-07-12 Online:2018-05-20 Published:2018-05-23

摘要/Abstract

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。

关键词: BST-2基因, 原核表达, 真核表达, 亚细胞定位, 山羊

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.

Key words: BST-2 gene (bone marrow stromal antigen 2 gene), prokaryotic expression, eukaryotic expression, subcellular localization, Capra hiracs

中图分类号:

S827

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): 702-706.

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]. , 2018, 30(5): 702-706.

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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

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