| 香蕉束顶病毒海口分离物CP基因的原核表达、纯化及其抗血清制备 |
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| 引用本文： | 余乃通,冯团诚,王健华,张雨良,刘志昕. 香蕉束顶病毒海口分离物CP基因的原核表达、纯化及其抗血清制备[J]. 热带作物学报, 2010, 31(5): 767-771. DOI: 10.3969/j.issn.1000-2561.2010.05.014 |
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| 作者姓名： | 余乃通 冯团诚 王健华 张雨良 刘志昕 |
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| 作者单位： | 1. 中国热带农业科学院热带生物技术研究所,农业部热带作物生物技术重点开放实验室,海南,海口,571101;海南大学环境与植物保护学院,海南,海口,570228
2. 中国热带农业科学院热带生物技术研究所,农业部热带作物生物技术重点开放实验室,海南,海口,571101 |
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| 基金项目： | 国家科技支撑计划项目，教育部/海南省教育厅联合资助博士点基金 |
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| 摘 要： | 用PCR方法从感染香蕉束顶病毒(Banana bunchy top virus,BBTV)的香蕉植株幼嫩假茎和叶片总DNA中扩增外壳蛋白(coat protein,CP)基因。回收目的片段,经XhoⅠ和BamHⅠ双酶切后与同样酶切的质粒载体pET30a(+)相连接,酶切验证及测序结果获得了含BBTV CP基因的重组质粒pET30a-CP。将重组质粒转化Codon plus表达菌,在25℃、0.4mmol/L IPTG条件下诱导6h或过夜,经12%SDS-PAGE电泳分析可知,该重组菌表达了一个与预期分子量大小一致的His-CP融合蛋白,约为30ku,且主要以包涵体的形式存在。多次洗涤包涵体后得到高纯度的融合蛋白,用其免疫家兔,获得BBTV CP蛋白抗血清。间接ELISA法测定抗血清效价大于51200。Western blot分析表明,抗血清能与融合蛋白特异性结合。
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| 关 键 词： | 香蕉束顶病毒 CP基因 原核表达 纯化 抗血清 |
Prokaryotic expression and Purification of CP gene of Banana Bunchy Top Virus HaiKou Isolate and Preparation of Antiserum Against its CP |
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| Yu Naitong,Feng Tuancheng,Wang Janhu,Zhang Yuliang and Liu Zhixin. Prokaryotic expression and Purification of CP gene of Banana Bunchy Top Virus HaiKou Isolate and Preparation of Antiserum Against its CP[J]. Chinese Journal of Tropical Crops, 2010, 31(5): 767-771. DOI: 10.3969/j.issn.1000-2561.2010.05.014 |
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| Authors: | Yu Naitong Feng Tuancheng Wang Janhu Zhang Yuliang Liu Zhixin |
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| Affiliation: | Key Laboratory for Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology, CATAS;1 Key Laboratory for Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology, CATAS 2 College of Environmemt and Plant Protection,Hainan University;Key Laboratory for Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology, CATAS;Key Laboratory for Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology, CATAS;Key Laboratory for Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology, CATAS |
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