| TeiR融合蛋白分离及多克隆抗体的制备 |
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| 引用本文： | 陈建秋,艾育芳,潘大仁,周以飞,潘菲,林彬辉,郭玉春.TeiR融合蛋白分离及多克隆抗体的制备[J].福建农林大学学报(自然科学版),2009,38(3). |
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| 作者姓名： | 陈建秋 艾育芳 潘大仁 周以飞 潘菲 林彬辉 郭玉春 |
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| 作者单位： | 1. 福建农林大学作物科学学院
2. 福建农林大学生命科学学院,福建,福州,350002 |
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| 基金项目： | 教育部科学技术研究重点项目，福建省自然科学基金，福建省教育厅科学技术重点项目，福建省发改委资助项目 |
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| 摘 要： | 提取睾丸酮丛毛单胞菌(Comamonas testosteroni)基因组DNA,PCR扩增teiR基因,将扩增产物克隆到pET28a中,经酶切验证后获得teiR基因表达载体pET28a-teiR.将重组质粒转化大肠杆菌(Escherichia coli)BL21,添加终浓度为1 mmol.L-1的IPTG进行诱导,提取细菌总蛋白进行蛋白的SDS-PAGE电泳.并根据融合蛋白特性,利用Ni-NTA Spin Column对TeiR融合蛋白进行分离纯化,经分离纯化后的重组蛋白在SDS-PAGE上呈现出单一条带.进一步以获得的TeiR融合蛋白为抗原,制备兔抗TeiR多克隆抗体,经ELISA测定该抗体的效价为1∶10000,该抗体与睾丸酮丛毛单胞菌天然TeiR蛋白反应良好,说明该抗体特异性良好.
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| 关 键 词： | 睾丸酮丛毛单胞菌 teiR SDS-PAGE 多克隆抗体 |
Separation of TeiR fusion protein and its multiclonal antibody preparation |
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| CHEN Jian-qiu,AI Yu-fang,PAN Da-ren,ZHOU Yi-fei,PAN Fei,LIN Bing-hui,GUO Yu-chun.Separation of TeiR fusion protein and its multiclonal antibody preparation[J].Journal of Fujian Agricultural and Forestry University,2009,38(3). |
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| Authors: | CHEN Jian-qiu AI Yu-fang PAN Da-ren ZHOU Yi-fei PAN Fei LIN Bing-hui GUO Yu-chun |
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| Institution: | CHEN Jian-qiu1,AI Yu-fang2,PAN Da-ren2,ZHOU Yi-fei1,PAN Fei2,LIN Bing-hui2,GUO Yu-chun1(1.College of Crop Science,2.College of Life Science,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China) |
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| Abstract: | The teiR gene was amplified by PCR from the genomic DNA of Comamonas testosteroni and then cloned into plasmid pET28a to construct recombinant plasmid pET28a-teiR.Plasmid pET28a-teiR was transformed into Escherichia coli BL21.The total cell protein of the host bacteria was extracted for SDS-PAGE after inducing with 0.001 mol·L-1 IPTG.TeiR fusion protein was purified using Ni-NTA Spin Column for multiclonal antibody preparation.The results of ELISA showed that the titer of the multiclonal antibody against the purified TeiR fusion protein was about 1∶10000.The antibody reacted well against the native TeiR protein from C.testosteroni. |
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| Keywords: | teiR SDS-PAGE |
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