Tools for manipulation of secondary metabolism pathways: rapid promoter replacements and gene deletions in Aspergillus nidulans.
Tools for manipulation of secondary metabolism pathways: rapid promoter replacements and gene deletions in Aspergillus nidulans.
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DOI：
10.1007/978-1-62703-122-6_10
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发表时间：
2012-01-01
期刊：
影响因子：
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通讯作者：
Oakley, Berl R
中科院分区：
文献类型：
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作者：
Oakley, C Elizabeth;Edgerton-Morgan, Heather;Oakley, Berl R
靶向基因缺失和启动子替换被证明是一种有价值的工具，可以唤醒和分析灰曲霉中沉默的次级代谢基因簇，随着操纵其他真菌基因组的分子遗传学方法的发展，它们可能在这些生物中同样有价值。在这里，我们描述了通过PCR构建DNA片段的程序，这些片段可用于快速和大规模地替换基因或启动子。我们还描述了a . nidulans的转化过程，使这些片段能够有效地引入目标菌株，从而在一次实验中可以替换许多基因或启动子。
Targeted gene deletions and promoter replacements are proving to be a valuable tool for awakening and analyzing silent secondary metabolism gene clusters in Aspergillus nidulans and, as molecular genetic methods for manipulating the genomes of other fungi are developed, they will likely be as valuable in those organisms. Here we describe procedures for constructing DNA fragments by PCR that can be used to replace genes or promoters quickly and on a large scale. We also describe transformation procedures for A. nidulans that allow these fragments to be introduced into target strains efficiently such that many genes or promoters can be replaced in a single experiment.