| 丝状真菌转化载体的改进 |
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| 引用本文： | 李继刚,陈永利,杨忠祥,郑建坡.丝状真菌转化载体的改进[J].河南农业科学,2013,42(5). |
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| 作者姓名： | 李继刚 陈永利 杨忠祥 郑建坡 |
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| 作者单位： | 河北大学生命科学学院,河北省微生物多样性研究与应用实验室,河北保定071002 |
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| 基金项目： | 河北省科技支撑计划项目，河北省重点实验室开放课题 |
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| 摘 要： | 为了改进农杆菌介导丝状真菌遗传转化用的Ti双元载体,将质粒pUCATPH上真菌trpC基因启动子驱动的潮霉素B抗性基因(hygB)表达盒转移到Ti质粒pPZP111上,构建成用于丝状真菌遗传转化的Ti载体pATZ。将植物表达载体pDL916上的草丁膦抗性bar基因重组到载体pKS-trpC上,构建成含真菌启动子、终止子调控的bar基因表达盒的中间载体pTrpCBar;之后将该表达盒转移到pPZP111载体上,得到以bar基因为筛选标记的真菌转化Ti质粒pTBZ。上述质粒载体经酶切鉴定和DNA测序证实构建正确,为丝状真菌的农杆菌介导遗传转化提供了新的工具。
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| 关 键 词： | 农杆菌介导转化 丝状真菌 转化载体 草丁膦抗性基因bar 潮霉素B抗性基因hygB 真菌启动子 |
Modification of Genetic Transformation Vectors for Filamentous Fungi |
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| LI Ji-gang
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CHEN Yong-li
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YANG Zhong-xiang
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ZHENG Jian-po.Modification of Genetic Transformation Vectors for Filamentous Fungi[J].Journal of Henan Agricultural Sciences,2013,42(5). |
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| Authors: | LI Ji-gang CHEN Yong-li YANG Zhong-xiang ZHENG Jian-po |
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| Abstract: | In order to improve the efficiency of Agrobacterium-mediated transformation of filamentous fungi(ATMT),a fungal promoter-driven hygromycin resistance gene(hygB) cassette of pUCATPH was transferred to Ti plasmid pPZP111,resulting in Ti binary vector pATZ.The glufosinate resistance gene(bar) of pDL916 was inserted to pKS-trpC to construct a new fungal promoter-driven bar gene expression cassette,which was recombined to pPZP111 to form a new Ti binary vector pTBZ.The recombinant binary Ti plasmids were confirmed by restriction enzyme digestion and DNA sequencing.The above vectors constructed may provide new tools for ATMT of filamentous fungi. |
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| Keywords: | Agrobacterium-mediated transformation(ATMT) filamentous fungi transformation vector glufosinate resistance gene bar hygromycin B resistance gene hygB fungal promoter |
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