| 极细链格孢菌peaT1基因在毕赤酵母中的表达与功能 分析 |
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| 引用本文： | 刘延锋,曾洪梅,玉山江,杨秀芬,毛建军,邱德文.极细链格孢菌peaT1基因在毕赤酵母中的表达与功能 分析[J].生物工程学报,2009,25(3):413-417. |
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| 作者姓名： | 刘延锋 曾洪梅 玉山江 杨秀芬 毛建军 邱德文 |
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| 作者单位： | 中国农业科学院植物保护研究所,植物病虫害生物学国家重点实验室,北京,100081 |
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| 基金项目： | 国家重点基础研究发展规划项目(973项目)(No.2003CB114204);;国家高技术研究发展计划(863计划)(No.2006AA10A210);;国家自然科学基金项目(No.30571252)资助~~ |
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| 摘 要： | 建立了在毕赤酵母(Pichiapastoris)中分泌表达PeaT1蛋白的技术。将来源于极细链格孢菌(Alternaria tenuissima)的基因peaT1亚克隆至酵母分泌型表达载体pPIC9K,构建了重组表达载体pPIC9K-peaT1,分别用SalI或BglII酶切线性化后电击转入毕赤酵母GS115菌株中,经MD平板筛选、PCR鉴定获得整合有外源基因的重组菌株。在α-Factor及AOX1基因启动子和终止信号的调控下,PeaT1在酵母中大量表达并分泌到胞外,SDS-PAGE检测表明表达蛋白的表观分子量约为35kD。表达蛋白上清稀释液能诱导烟草产生对TMV的抗性,其枯斑数抑制率可达到30.37%。每升表达上清液经超滤浓缩和离子交换层析可纯化目的蛋白16.13mg,该纯化蛋白能显著地促进小麦幼苗的生长。
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| 关 键 词： | peaT1 极细链格孢菌 毕赤酵母 |
| 收稿时间： | 2008/3/27 0:00:00 |
Expression H113of peaT1 gene from Alternaria tenuissima in Pichia pastoris and its function |
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| Yanfeng Liu,Hongmei Zeng,Shanjiang Yu,Xiufen Yang,Jianjun Mao and Dewen Qiu.Expression H113of peaT1 gene from Alternaria tenuissima in Pichia pastoris and its function[J].Chinese Journal of Biotechnology,2009,25(3):413-417. |
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| Authors: | Yanfeng Liu Hongmei Zeng Shanjiang Yu Xiufen Yang Jianjun Mao and Dewen Qiu |
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| Institution: | State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100081, China;State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100081, China;State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100081, China;State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100081, China;State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100081, China;State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100081, China |
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| Abstract: | |
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| Keywords: | peaT1 Alternaria tenuissima Pichia pastoris |
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