丝状真菌的基因工程:粗神经孢子菌转化酶基因的克隆。

M Carú
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引用次数: 0

摘要

将稻瘟病菌转化酶野生型基因克隆到酵母载体YRp7中。该重组质粒是通过酿酒葡萄球菌突变株的功能互补获得的。分离得到的重组质粒pNC2携带一段7.6 Kb的稻瘟菌BamHI DNA片段。克隆的DNA与油菜基因组DNA杂交,将油菜Inv-转化为Inv+转化酶突变体。N. crassa的Inv-到Inv+的变换似乎至少需要两个不同的积分事件。其中一个涉及与inv基因座紧密相连的整合,另一个显然涉及克隆DNA在与inv基因座不同的基因组位点的整合。
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[Genetic engineering in filamentous fungi: cloning of the invertase gene from Neurospora crassa].

The invertase wild type gene of N. crassa was cloned into the YRp7 yeast vector. This recombinant plasmid was selected by functional complementation of an invertaseless mutant strain of S. cerevisiae. The isolated recombinant plasmid (named pNC2) carries a 7.6 Kb BamHI DNA fragment from N. crassa. The cloned DNA hybridized with the N. crassa genomic DNA and transformed an invertase mutant of N. crassa Inv- to Inv+. Transformation of N. crassa Inv- to Inv+ seems to take at least two different integration events. One of them involves an integration closely linked to inv locus, and the other one apparently involves an integration of cloned DNA at a genomic site different that the inv locus.

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