应用努尔塞乌霉素乙酰转移酶基因(nat1)作为丝状真菌转化的显性标记

U. Kück, Birgit Hoff, Lehrstuhl für Allgemeine
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引用次数: 78

摘要

在此,我们构建了两个转化载体pD-NAT1和pG-NAT1,它们分别携带编码诺斯基丙氨酸乙酰转移酶的nat1基因。nat1基因在细粒曲霉trpC启动子的调控下表达,可以作为丝状真菌dna介导转化的显性耐药标记物。通过转化同源子囊菌大孢子索达菌(Sordaria macrospora)和产β-内酰胺的Acremonium chrysogenum,证明了这两种载体的成功应用。对于真菌和两种载体,每10µg质粒DNA的转化频率在10到40个转化子之间。
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Application of the nourseothricin acetyltransferase gene (nat1) as dominant marker for the transformation of filamentous fungi
Here, we report the construction of two transformation vectors, pD-NAT1 and pG-NAT1, carrying the nat1 gene encoding the nourseothricin acetyltransferase. The nat1 gene is expressed under the control of the Aspergillus nidulans trpC promoter and thus can be used as a dominant drug-resistance marker for the DNA-mediated transformation of filamentous fungi. The successful application of both vectors was demonstrated by transforming the homothallic ascomycete Sordaria macrospora as well as the β-lactam producer Acremonium chrysogenum. For both fungi and for both vectors, transformation frequencies were between 10 and 40 transformants per 10 µg of plasmid DNA.
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