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| 新加坡石斑鱼虹彩病毒ORF162蛋白的表达、纯化及抗体制备 | |
| 引用本文： | 夏立群,梁海鹰,江韶英,张尧清.新加坡石斑鱼虹彩病毒ORF162蛋白的表达、纯化及抗体制备[J].广东海洋大学学报,2010,30(1):59-64. |
| 作者姓名： | 夏立群 梁海鹰 江韶英 张尧清 |
| 作者单位： | 1. 广东海洋大学水产学院,广东,湛江,524025;中山大学生命科学学院,广东,广州,510275 2. 广东海洋大学水产学院,广东,湛江,524025;广东省水产经济动物病原生物学及流行病学重点实验室,广东,湛江,524088 3. 广东海洋大学水产学院,广东,湛江,524025 |
| 摘 要： | 将新加坡石斑鱼虹彩病毒(Singapore grouper iridovirus,SGIV)的ORF162的开放式阅读框插入pET-32a表达载体T7启动子控制下的6-His·Tag编码基因上游,构建SGIVORF162原核表达质粒pET-ORF162。表达质粒转化入大肠杆菌BL21(DE3)菌株,经IPTG诱导,成功表达SGIV ORF162融合蛋白。对IPTG浓度、诱导温度、诱导时间等诱导表达条件进行优化后,确定在0.7mmol/LIPTG、16℃条件下诱导14h时可溶性SGIV ORF162重组蛋白占重组蛋白总量的95%。经镍琼脂糖凝胶纯化,获得纯度为90%以上的SGIV ORF162蛋白。用纯化的SGIV ORF162蛋白免疫小鼠,获得高效特异的SGIV ORF162多克隆抗体。 |
| 关 键 词： | 新加坡石斑鱼虹彩病毒 原核表达 多克隆抗体 |
Expression,Purification and Antibody Preparation of Singapore Grouper Iridovirus ORF162 |
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| XIA Li-qun,LIANG Hai-ying,JIANG Shao-ying,ZHANG Rao-qing.Expression,Purification and Antibody Preparation of Singapore Grouper Iridovirus ORF162[J].Journal of Zhanjiang Ocean University,2010,30(1):59-64. | |
| Authors: | XIA Li-qun LIANG Hai-ying JIANG Shao-ying ZHANG Rao-qing |
| Institution: | XIA Li-qun 1,2,LIANG Hai-ying 1,3,JIANG Shao-ying 1,ZHANG Rao-qing 1 (1.College of Fishery,Guangdong Ocean University,Zhanjiang 524025,China,2.School of Life Science,Sun Yat-sen University,3.Guangdong Provincial Key Laboratory of Pathogenic Biology , Epidemiology for Aquatic Economic Animals,China) |
| Abstract: | Full length of SGIV ORF162 was inserted into the prokaryotic vector pET-32a fused with the down stream 6-His?Tag.The expression vector was transformed into the E.coli BL21 (DE3) strain and SGIV ORF162 fusion protein was successfully expressed.The optimum condition for the expression of soluble SGIV ORF162 fusion protein was 0.7 mmol/L IPTG,16 ℃,14h.The fusion protein were purified by Ni-IDA His?Bind affinity column and the purity of recombinant SGIV ORF162 protein was above 90%.The polyclonal antibody is prepared by immunizing mice with purified SGIV ORF162 protein and proved to be specific against SGIV ORF162 protein.The purified SGIV ORF162 fusion protein and the polyclonal antibody can be used for further functional and structural studies. |
| Keywords: | Singapore grouper iridovirus (SGIV) prokaryotic expression polyclonal antibody |
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