| 益母草抗菌蛋白LJAMP2的表达及抗菌活性分析 |
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| 引用本文： | 郭绍辉,杨永柱,王晓雯,谢成建,杨星勇.益母草抗菌蛋白LJAMP2的表达及抗菌活性分析[J].农业生物技术学报,2010,18(2):254-259. |
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| 作者姓名： | 郭绍辉 杨永柱 王晓雯 谢成建 杨星勇 |
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| 作者单位： | 1. 三峡库区生态环境教育部重点实验室,西南大学生命科学学院,重庆,400716
2. 重庆市九龙坡区农业局,重庆,400010 |
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| 基金项目： | 国家自然科学基金，重庆市科委自然科学基金 |
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| 摘 要： | LJAMP2是一种分离自益母草(Leonurus japonicus Houtt.)种子的非专一性脂转移蛋白类抗菌蛋白.将编码成熟的LJAMP2蛋白的基冈(GenBank Accession No.AY971513)插入到巴斯德毕赤酵母(Pichia pas-toris)表达载体pPIC9K中;重组质粒pPIC9K-LJAMP2经线性化后,用电转化法导入毕赤酵母菌株GS115(his-/mut+)中.转化子用含1.75 mg/mL G418的YPD平板筛选获得多拷贝Mut+转化子.将1%甲醇诱导LJAMP2表达后的发酵液,经进一步浓缩、凝胶过滤脱盐、CM FF阳离子交换色谱和反相HPLC进行纯化,获得纯度90%以上的LJAMP2菌株诱导24 h即有LJAMP2的表达,诱导168 h表达量达到最高;表达的LJAMP2蛋白对黑曲霉(Aspergillus niger)、玉米小斑病菌(Bipolaris maydis)、镰刀菌(Fusarium graminearum)真菌表现出抑制活性.
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| 关 键 词： | 益母草 抗菌蛋白 毕赤酵母 分泌表达 |
Purification and Antifungal Activities of an Antimicrobial Protein LJAMP2 Expressed in Pichia pastoris |
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| Guo Shaohui,Yang Yongzhu,Wang Xiaowen,Xie Chengjian,Yang Xingyong.Purification and Antifungal Activities of an Antimicrobial Protein LJAMP2 Expressed in Pichia pastoris[J].Journal of Agricultural Biotechnology,2010,18(2):254-259. |
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| Authors: | Guo Shaohui Yang Yongzhu Wang Xiaowen Xie Chengjian Yang Xingyong |
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| Institution: | Guo Shaohui1 Yang Yongzhu2 Wang Xiaowen1 Xie Chengjian1 Yang Xingyong1 1 Key Laboratory of Eco-Environments in Three Gorges Reservoir Region,Ministry of Education,School of Life Science,Southwest University,Chongqing,400716,2 The Agricultural Bureau of Jiulongpo Area in Chongqing,400010 |
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| Abstract: | LJAMP2, a member ofnonspecific lipid transfer protein, isolated from motherwort seeds, displays antimicrobial activity against fungi.The LJAMP2 gene was cloned into the pPIC9K vector and transformed into Pichia pastoris usingelectrotransformation.A transformant with a high copynumber of the gene integrated into the chromosome was obtained by YPD plates which contained 1.75 mg/mL G418.The active LJAMP2 was secreted into the culture medium under the induction of 1% methanol and purified more than 90% purity f... |
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| Keywords: | Motherwort seed Antimicrobial protein Yeast Expression |
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