| 甘蔗花叶病毒CI蛋白在玉米韧皮部细胞中的免疫定位 |
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| 引用本文： | 秦艳红,刘冬梅,周涛,范在丰.甘蔗花叶病毒CI蛋白在玉米韧皮部细胞中的免疫定位[J].植物保护学报,2009,36(3):234-238. |
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| 作者姓名： | 秦艳红 刘冬梅 周涛 范在丰 |
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| 作者单位： | 1. 中国农业大学植物病理学系,农业生物技术国家重点实验室,北京,100193;河南省农业科学院植物保护研究所生物技术组,郑州,450002
2. 中国农业大学植物病理学系,农业生物技术国家重点实验室,北京,100193 |
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| 基金项目： | 农业生物技术国家重点实验室开放(青年)课题(2008SKLAB08-03), 国家自然科学基金(30571213) |
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| 摘 要： | 为探讨甘蔗花叶病毒(Sugarcane mosaic virus,SCMV)CI基因的功能,对CI蛋白进行了免疫定位.利用RT-PCR方法,扩增甘蔗花叶病毒北京株系(SCMV Beijing isolate,SCMV-BJ)CI基因部分片段,将其连接到表达载体pET-28a上,重组质粒pETCIF1导入大肠杆菌BL21(DE3)pLysS中.SDS-PAGE分析表明,经IPTG诱导,CI基因获得了高效表达.将融合蛋白纯化后免疫兔子获得特异性较高的抗血清,利用该抗血清,对健康玉米和受病毒侵染的玉米茎、叶脉和叶片进行免疫金标记,在茎和叶脉韧皮部筛管的细胞壁处有金颗粒.
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| 关 键 词： | 甘蔗花叶病毒 CI基因 原核表达 免疫金定位 |
| 收稿时间： | 2008/12/9 0:00:00 |
Immuno-localization of the CI protein of Sugarcane mosaic virus in the phloem cells of maize |
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| Qin Yanhong,Liu Dongmei,Zhou Tao and Fan Zaifeng.Immuno-localization of the CI protein of Sugarcane mosaic virus in the phloem cells of maize[J].Acta Phytophylacica Sinica,2009,36(3):234-238. |
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| Authors: | Qin Yanhong Liu Dongmei Zhou Tao and Fan Zaifeng |
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| Institution: | Department of Plant Pathology, and State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing 100193, China; Institute of Plant Protection, Henan Academy of Agricultural Sciences, Zhengzhou 450002, Henan Province, China;Department of Plant Pathology, and State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing 100193, China;Department of Plant Pathology, and State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing 100193, China;Department of Plant Pathology, and State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing 100193, China |
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| Abstract: | In order to explore the function of Sugarcane mosaic virus CI gene, the immuno-localization of CI protein was investigated. Partial fragment of CI gene of SCMV Beijing isolate was amplified by RT-PCR, and cloned into the expression vector pET-28a. The recombinant plasmid was transformed into E.coli BL21(DE3)pLysS. The result of SDS-PAGE analysis showed that CI gene was highly expressed by IPTG induction. The highly specific antiserum was produced when rabbit was immunized with purified fusion protein. The localization of the CI protein in maize cells of stem, vein and leaf tissues from SCMV-BJ-infected or healthy maize plants were examined by immuno-gold labeling technique,and the results indicated that this protein was localized in the phloem cell wall of SCMV-infected stems and veins. |
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| Keywords: | Sugarcane mosaic virus CI gene prokaryotic expression immuno-gold labeling localization |
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