| 稻瘟病菌一假定几丁质酶在毕赤酵母中的表达 |
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| 引用本文： | 周洁,陈鑫,郑祥梓,林成增,王宗华,鲁国东. 稻瘟病菌一假定几丁质酶在毕赤酵母中的表达[J]. 福建农林大学学报(自然科学版), 2009, 38(6) |
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| 作者姓名： | 周洁 陈鑫 郑祥梓 林成增 王宗华 鲁国东 |
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| 作者单位： | 1. 福建农林大学生命科学学院,福建,福州,350002
2. 福建农林大学植物保护学院,福建,福州,350002 |
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| 基金项目： | 国家自然科学基金，福建省自然科学基金 |
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| 摘 要： | 本研究从稻瘟病菌菌株Guy11中提取总RNA,利用RT-PCR方法扩增到一假定几丁质酶基因MGG_08054.6的cDNA序列,将其亚克隆至带有His-tag标签的酵母表达载体pPIC3.5K中,构成重组质粒pPIC3.5K-ws,电击转化至毕赤酵母菌菌株GS115,通过MD-MM平板筛选及PCR验证获得重组酵母转化子GS115-PICH-ws;阳性转化子在甲醇诱导下,成功分泌出重组蛋白.SDS-PAGE分析表明,重组蛋白分子质量为48 ku,与其理论计算值基本相符;Ni-NTA法分离纯化后的重组蛋白经DNS法测定具有几丁质酶活性.
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| 关 键 词： | 稻瘟病菌 几丁质酶 毕赤酵母 表达 |
Expression of a putative Magnaporthe oryzae chitinase in Pichia pastoris |
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| ZHOU Jie,CHEN Xin,ZHENG Xiang-zi,LIN Cheng-zeng,WANG Zong-hua,LU Guo-dong. Expression of a putative Magnaporthe oryzae chitinase in Pichia pastoris[J]. Journal of Fujian Agricultural and Forestry University, 2009, 38(6) |
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| Authors: | ZHOU Jie CHEN Xin ZHENG Xiang-zi LIN Cheng-zeng WANG Zong-hua LU Guo-dong |
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| Affiliation: | ZHOU Jie1,CHEN Xin1,ZHENG Xiang-zi1,LIN Cheng-zeng1,WANG Zong-hua1,LU Guo-dong2(1.College of Life Science,2.College of Plant Protection,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China) |
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| Abstract: | Chitinase may participate in regulating cell wall dynamics,mediating the growth and colonization of fungi.In this study,cDNA of a putative chitinase gene(MGG_08054.6) was amplified by RT-PCR from the total RNA of Magnaporthe oryzae strain Guy 11.It was then subcloned into the Pichia expression vector pPIC3.5K with a RGS-His-tag,generating a recombinant plasmid,pPIC 3.5K-ws.To express the fusion protein,linearized pPIC3.5K-ws was transformed into P.pastoris strain GS115 by electroporation.Positive recombinan... |
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| Keywords: | Magnaporthe oryzae chitinase Pichia pastoris expression |
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