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用于玉米黑粉菌基因操作的穿梭载体。

Shuttle vectors for genetic manipulations in Ustilago maydis.

作者信息

Kojic M, Holloman W K

机构信息

Department of Microbiology and Immunology, Cornell University Weill Medical College, New York, NY 10021, USA.

出版信息

Can J Microbiol. 2000 Apr;46(4):333-8. doi: 10.1139/w00-002.

Abstract

Shuttle vectors with new or improved features were constructed to enable facile genetic manipulations in the plant pathogen Ustilago maydis. Sets of plasmids selectable in media containing geneticin, carboxin, nourseothricin, or hygromycin, able to replicate autonomously, to transform U. maydis by integration, and to express foreign genes under control of the homologous glyceraldehyde-3-phosphate dehydrogenase promoter, were built upon a common pUC19 vector backbone. This permits a large number of choices for a cloning site, blue/white screening for recombinant plasmids, rapid transfer of a cloned DNA fragment between plasmids, and choice of several dominant drug-resistance markers for selection in U. maydis.

摘要

构建了具有新特征或改进特征的穿梭载体,以便在植物病原菌玉米黑粉菌中进行便捷的基因操作。基于常见的pUC19载体骨架构建了一组质粒,这些质粒在含有遗传霉素、羧苄青霉素、诺尔斯菌素或潮霉素的培养基中可选择,能够自主复制,通过整合转化玉米黑粉菌,并在同源甘油醛-3-磷酸脱氢酶启动子的控制下表达外源基因。这为克隆位点提供了大量选择,可对重组质粒进行蓝/白筛选,在质粒之间快速转移克隆的DNA片段,并可选择几种显性抗药标记用于玉米黑粉菌的选择。

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