npt Ⅱ基因真菌表达载体的构建及在苹果炭疽叶枯病菌遗传转化中的应用
Construction of the Fungus Expression Vector of npt Ⅱ Gene and Applying to the Genetic Transformation in Glomerella cingulata
查看参考文献14篇
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文摘
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本研究旨在构建1个适用于苹果炭疽叶枯病菌(Glomerella cingulata)的遗传转化载体,为系统研究该病原菌基因的功能提供便利。通过基因工程及分子生物学技术,将三磷酸甘油醛脱氢酶(GAPDH)基因启动子Pgap和新霉素磷酸转移酶基因(npt Ⅱ)连结于质粒pCambia0380中。构建的载体pGapneoRl导入根癌农杆菌LBA4404后,通过农杆菌介导的转化技术(ATMT),成功将npt Ⅱ基因盒整合到苹果炭疽叶枯病菌基因组中。Southern blot分析结果表明,随机挑取的转化子T-DNA均以单拷贝插入到苹果炭疽叶枯病菌基因组中,表明该载体适合于苹果炭疽叶枯病菌的遗传转化。此外,该载体也可用于构建苹果炭疽叶枯病菌的目标基因的回补载体、超表达载体和荧光融合蛋白表达载体。 |
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其他语种文摘
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This study is aimed to construct one vector that applys to the genetic transformation of Glomerella cingulata, to facilitate the research in the gene function of the fungus. Pgap promoter of the gene encoding glyc-eraldehyde-3-phosphate dehydrogenase (GAPDH) and neomycinphosphotransferase gene (npt Ⅱ) were ligated into the plasmid pCambia0380, respectively, using biology and genetic engineering theory and technology. The npt Ⅱgene cassette was introduced to the genome of G. cingulata via Agrobacterium tumefaciens-mediated transformation (ATMT), after the obtained plasmid pGapneoRl was transformed to Agrobacterium tumefaciens. The Southern blot analysis revealed that all T-DNAs of the randomly transformants picked were a single copy in the genomes of G. cingulata, and showed that this plasmid would be suitable to genetic transformation of Glomerella cingulata. Moreover, this vector can be used to construct the restoration vector, overexpressed vector and fluorescent protein fused vector. |
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来源
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基因组学与应用生物学
,2015,34(10):2156-2160 【核心库】
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DOI
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10.13417/j.gab.034.002156
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关键词
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炭疽病
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新霉素磷酸转移酶基因
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农杆菌
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启动子
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地址
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中国农业科学院果树研究所, 兴城, 125100
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语种
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中文 |
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文献类型
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研究性论文 |
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ISSN
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1674-568X |
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学科
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分子生物学;园艺 |
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基金
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现代苹果产业技术体系建设项目
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文献收藏号
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CSCD:5557851
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