Genetic manipulation of Aspergillus nidulans:: heterokaryons and diploids for dominance, complementation and haploidization analyses
Genetic manipulation of Aspergillus nidulans:: heterokaryons and diploids for dominance, complementation and haploidization analyses
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DOI：
10.1038/nprot.2007.113
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发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
Hynes, Michael J.
中科院分区：
文献类型：
--
作者：
Todd, Richard B.;Davis, Meryl A.;Hynes, Michael J.
单倍体微生物真核生物构巢曲霉是一个强大的遗传系统，它允许分析广泛的生物现象。除了常规的单代减数分裂后代分析外，拟性分析提供了一种快速、方便的遗传分析方法。我们描述了A.用于遗传分析以确定优势和进行互补试验的nidulans异核体和二倍体。我们还描述了通过携带标记染色体的二倍体的单倍体化将突变快速映射到染色体。平衡异核体可在10天内建立，二倍体可在2-3周内构建。利用平衡异核体或二倍体进行的显性试验和互补试验可在2-3天内完成。杂合二倍体的单倍体化分析可以在10天内实现。这些协议应适用于相关的阿氏菌和青霉菌，缺乏一个已知的减数分裂周期。
The haploid microbial eukaryote Aspergillus nidulans is a powerful genetic system, which allows analysis of a broad range of biological phenomena. In addition to conventional analysis of meiotic progeny in a single generation, parasexual analysis affords a rapid and convenient method for genetic analysis. We describe the construction of A. nidulans heterokaryons and diploids for use in genetic analysis to determine dominance and conduct complementation tests. We also describe the rapid mapping of mutations to chromosomes by haploidization of diploids carrying marked chromosomes. Balanced heterokaryons may be established within 10 days and diploids may be constructed in 2-3 weeks. Dominance tests and complementation tests using balanced heterokaryons or diploids may be completed in 2-3 days. Haploidization analysis of heterozygous diploids can be achieved within 10 days. These protocols should be adaptable for use in related Aspergilli and Penicillia, which lack a known meiotic cycle.