| 水稻核黄素合酶基因的克隆、表达及多克隆抗体的制备 |
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| 引用本文： | 孙力军,刘会敏,任海英,吴廷全,宋韬,董汉松.水稻核黄素合酶基因的克隆、表达及多克隆抗体的制备[J].江苏农业学报,2010,26(2). |
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| 作者姓名： | 孙力军 刘会敏 任海英 吴廷全 宋韬 董汉松 |
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| 作者单位： | 南京农业大学植物保护学院植物病理学系,农业部作物病虫害监测与防控重点开放实验室,江苏,南京,210095 |
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| 基金项目： | 国家杰出青年科学基金(30525088) |
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| 摘 要： | 核黄素在生物体内发挥着重要的作用,核黄素合酶(RS)是核黄素合成过程中的一个关键酶。本研究克隆了水稻核黄素合酶基因(OsRS),并将其在Escherichia coli中高效表达。表达产物(包涵体),先用6 mol/L盐酸胍将其溶解,再用8 mol/L尿素透析,将获得的蛋白免疫家兔制备了多克隆抗体。并将抗体与原核表达的OsRS和提取的水稻叶组织总蛋白进行Western blot分析,确定所制备的多克隆抗体具有较好的免疫反应性,为深入解析水稻中核黄素合酶的催化活性和功能奠定了基础。
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| 关 键 词： | 水稻 核黄素合酶 包涵体 多克隆抗体 |
Cloning and Expression of Riboflavin Synthase Gene in Rice and Preparation of Polyclonal Antibody against Riboflavin Synthase |
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| SUN Li-jun,LIU Hui-min,REN Hai-ying,WU Ting-quan,SONG Tao,DONG Han-song.Cloning and Expression of Riboflavin Synthase Gene in Rice and Preparation of Polyclonal Antibody against Riboflavin Synthase[J].Jiangsu Journal of Agricultural Sciences,2010,26(2). |
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| Authors: | SUN Li-jun LIU Hui-min REN Hai-ying WU Ting-quan SONG Tao DONG Han-song |
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| Institution: | College of Plant Protection/a>;Nanjing Agricultural University/a>;Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects/a>;Ministry of Agriculture/a>;Nanjing 210095/a>;China |
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| Abstract: | Riboflavin synthase(RS) catalyzes the committed step in biosynthesis of riboflavin,which plays an important role in organisms.Here,riboflavin synthase gene in rice(OsRS) was cloned and expressed in Escherichia coli with high efficiency.Since the product of expressed OsRS occurred in the form of inclusion body,the protein was dissolved with 6 mol/L guanidine hydrochloride solution and then was dialysed with 8 mol/L urea solution.The rabbit anti-OsRS polyclonal antibody was prepared.Furthermore,antigen-specif... |
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| Keywords: | rice riboflavin synthase inclusion body polyclonal antibody |
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