| 高粱花叶病毒Hcpro、CP和PIPO基因酵母双杂交载体构建及自激活验证 |
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| 引用本文： | 张雨良,黄启星,张树珍,王健华,余乃通,刘志昕.高粱花叶病毒Hcpro、CP和PIPO基因酵母双杂交载体构建及自激活验证[J].热带作物学报,2012,33(12):2252-2256. |
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| 作者姓名： | 张雨良 黄启星 张树珍 王健华 余乃通 刘志昕 |
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| 作者单位： | 中国热带农业科学院热带生物技术研究所,海南海口 571101;农业部热带作物生物学与遗传资源利用重点实验室,海南海口571101 |
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| 基金项目： | 中央级公益性科研院所基本科研业务专项（ITBB110303）；现代农业产业体系建设专项资金（nycytx-24）；国家科技支撑计划课题（2007BAD48B01）。 |
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| 摘 要： | 采用同源克隆技术获得侵染甘蔗高粱花叶病毒(Sorghum mosaic virus,SrMV)的Hcpro、CP、PIPO基因片段.将该基因片段连接至pGBKT7载体,分别构建酵母双杂交诱饵载体pGBKTT-Hcpro、pGBKT7-CP和pGBKTT-PIPO,经PCR、酶切及测序鉴定,证明重组诱饵载体构建成功后,将重组质粒导入Y2H Gold酵母菌株,检测其表达产物对酵母细胞有无毒性及对报告基因有无激活作用.结果表明,含重组诱饵质粒的酵母菌株在SD/-Trp营养缺陷平板上生长良好,表明重组诱饵质粒表达产物对酵母细胞无毒性.含重组诱饵质粒的酵母菌株在SD/-Leu、SD/-Trp/-Ade和SD/-Trp/-His营养缺陷平板上不能生长,表明重组诱饵质粒表达产物对ADE2、HIS3报告基因无自激活作用.研究结果为下一步利用酵母双杂交系统检测与高粱花叶病毒Hcpro、CP、PIPO蛋白相互作用的甘蔗蛋白提供基础.
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| 关 键 词： | 高粱花叶病毒 Hcpro CP PIPO 酵母双杂交 诱饵载体 |
Construction of Bait Vectors Containing Hcpro, CP and PIPO Genes from Sorghum mosaic Virus in Yeast Two-hybrid System and Identification of Their Autonomous Activation |
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| ZHANG Yulian,HUANG Qixing,ZHANG Shuzhen,WANG Jianhu,YU Naitong and LIU Zhixin.Construction of Bait Vectors Containing Hcpro, CP and PIPO Genes from Sorghum mosaic Virus in Yeast Two-hybrid System and Identification of Their Autonomous Activation[J].Chinese Journal of Tropical Crops,2012,33(12):2252-2256. |
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| Authors: | ZHANG Yulian HUANG Qixing ZHANG Shuzhen WANG Jianhu YU Naitong and LIU Zhixin |
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| Institution: | 1 Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences
2 Key Laboratory of Biology and Genetic Resources of Torpical Crops, Ministry of Agriculture;1 Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences
2 Key Laboratory of Biology and Genetic Resources of Torpical Crops, Ministry of Agriculture;1 Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences
2 Key Laboratory of Biology and Genetic Resources of Torpical Crops, Ministry of Agriculture;1 Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences
2 Key Laboratory of Biology and Genetic Resources of Torpical Crops, Ministry of Agriculture;1 Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences
2 Key Laboratory of Biology and Genetic Resources of Torpical Crops, Ministry of Agriculture;1 Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences
2 Key Laboratory of Biology and Genetic Resources of Torpical Crops, Ministry of Agriculture |
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| Abstract: | |
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| Keywords: | Sorghum mosaic virus Hcpro CP PIPO Yeast two-hybrid Bait vector |
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