| 甘蓝型油菜植物防御素基因的克隆与原核表达 |
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| 引用本文： | 王轶,罗勤,吴俊,郭经宇,杨毅,李旭锋.甘蓝型油菜植物防御素基因的克隆与原核表达[J].四川大学学报(自然科学版),2005,42(4):831-834. |
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| 作者姓名： | 王轶 罗勤 吴俊 郭经宇 杨毅 李旭锋 |
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| 作者单位： | 四川大学生命科学学院植物遗传实验室,成都,610064 |
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| 摘 要： | 根据Genbank上已知的植物防御素基因设计引物，以甘蓝型油菜中的品种”蜀杂九号”种子总RNA反转录成的eDNA为模板，进行PCR扩增，获得了243bp的片段．将该片段回收。连接到pMD18-T载体测序，将测序片段经酶切回收后克隆到GTK表达载体中，在0．1mmol IPTG诱导下表达出与理论值相符的34kD的融合蛋白条带，用GST单克隆抗体做第一抗体进行Western-blot检测，获得阳性结果，这为甘蓝型油菜植物防御素基因功能的进一步研究提供了基础．
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| 关 键 词： | 甘蓝型油菜 植物防御素 融合蛋白 原核表达 |
| 文章编号： | 0490-6756（2005）04-0831-04 |
| 收稿时间： | 2005-01-27 |
| 修稿时间： | 2005-01-27 |
Gene Cloning and Prokaryotic Expression of Brassica napus Plant Defensin |
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| WANG Yi,LUO Qin,WU Jun,GUO Jing-yu,YANG Yi,LI Xu-feng.Gene Cloning and Prokaryotic Expression of Brassica napus Plant Defensin[J].Journal of Sichuan University (Natural Science Edition),2005,42(4):831-834. |
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| Authors: | WANG Yi LUO Qin WU Jun GUO Jing-yu YANG Yi LI Xu-feng |
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| Abstract: | Design a couple of primers according to the known sequence of plant defensins gene from Genbank.Extract the total RNA from Shuza No. 9 of Brassica napus seeds and then revert to cDNA. The gene was amplified by PCR and cloned into pMD18-T vecter , then was digested and inserted into the prokaryotic expression GTK vector. Protein was expressed under the induction of IPTG, and a protein band with a relative molecular weight of 34 kid was obtained and was recognized by GST antibody. This should provide a powerful tool for investigating the function of Brassica napus plant defensin. |
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| Keywords: | Brassica Napus plant defensins fusion protein prokaryotic expression |
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