| 小麦黄花叶病毒P1蛋白原核表达、抗血清制备及其检测 |
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| 引用本文： | 向荣,孙丽英,孙炳剑,陈剑平. 小麦黄花叶病毒P1蛋白原核表达、抗血清制备及其检测[J]. 浙江农业学报, 2011, 23(2): 0 |
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| 作者姓名： | 向荣 孙丽英 孙炳剑 陈剑平 |
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| 作者单位： | 1.浙江师范大学 化学与生命科学学院,浙江 金华 321000；2浙江省农业科学院 病毒与生物技术研究所,浙江省植物有害生物防控国家重点实验室培育基地,农业部植物保护与生物技术重点实验室,浙江省植物病毒重点实验室,浙江 杭州 310021 |
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| 基金项目： | 国家小麦产业技术体系项目，农业部小麦转基因项目 |
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| 摘 要： | 用RT-PCR方法从感染小麦黄花叶病毒(Wheat yellow mosaic virus,WYMV)扬州分离物的小麦叶片中扩增获得该病毒P1基因的全长cDNA片段,并进行了序列分析。结果表明，P1基因编码区含有765个核苷酸,编码1个由255氨基酸组成分子量为28.2 kDa的蛋白。将P1基因亚克隆到原核表达载体pGEX-6P-1中构建重组原核表达载体pGEX 6P-1-P1,经IPTG诱导,P1蛋白在大肠杆菌BL21(DE3)中表达GST-P1的融合蛋白,纯化并制备了P1蛋白的兔抗血清。利用此抗血清,可以检测WYMV病叶中的P1蛋白,但与提纯的WYMV粒子没有血清学反应,表明P1没有参与WYMV粒子的包装,是一种非结构蛋白。
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| 关 键 词： | 小麦黄花叶病毒 P1基因 克隆 原核表达 抗血清制备 血清学检测 |
Detection of P1 protein encoded by Wheat yellow mosaic virus RNA2, using polyclonal antibody raised against prokaryotic expression P1 fusion protein |
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| XIANG Rong,SUN Li-ying,SUN Bing-jian,CHEN Jian-ping. Detection of P1 protein encoded by Wheat yellow mosaic virus RNA2, using polyclonal antibody raised against prokaryotic expression P1 fusion protein[J]. Acta Agriculturae Zhejiangensis, 2011, 23(2): 0 |
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| Authors: | XIANG Rong SUN Li-ying SUN Bing-jian CHEN Jian-ping |
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| Affiliation: | 1College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321000, China; 2State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, MOA Key Laboratory for Plant Protection and Biotechnology, Zhejiang Provincial Key Laboratory of Plant Virology, Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China |
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| Abstract: | The complete cDNA of Wheat yellow mosaic virus(WYMV)P1 gene was obtained by RT-PCR from infected wheat leaves.The sequence analysis showed that P1 gene contained 765 nt,encoding a protein of 255 amino acids(aa) with estimated molecular weight of 28.2 kDa.The fragment of P1 ORF was cloned into prokaryotic expression vector pGEX-6P-1.Subsequently,IPTG-induced GST-P1 fusion protein was purified from E.coli BL21(DE3).The polyclonal antiserum of WYMV P1 was generated in rabbits by immunizing with purified GST-P1... |
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| Keywords: | Wheat yellow mosaic virus P1 gene cloning prokayotic expression polyconal antiserum immunoreaction |
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