Construction of minimally-sheltered knockouts mutants of Neurospora crassa

Fungal Genetics Reports Pub Date : 2005-12-01 DOI:10.4148/1941-4765.1123

R. L. Metzenberg

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引用次数: 2

Abstract

I have been developing strains which I think will make it possible to produce "minimally-sheltered knockouts" of essential genes routinely. They could equally well be called "self-adjusting knockdowns" of essential genes. Such minimally-sheltered knockouts could give results in microarray analysis that would be less subject to artifact than results with heterokaryons or with homokaryons sheltered with an inducible wild-type allele of the gene in question. The method addresses two questions about each cloned gene, henceforth and collectively called "your favorite gene" (yfg). yfg should be one which does not pass successfully through a cross as a hygromycin-resistant knockout. (1) Is yfg a dispensable gene? (2) If yfg is found to be essential, what is the "terminal phenotype" of the yfgΔ knockout mutation? By "terminal phenotype," I mean that of the heterokaryon between yfgΔ and a partial diploid, yfg+/Δ, in which the partial diploid is the minimum proportion that allows somewhat suboptimal growth. Creative Commons License This work is licensed under a Creative Commons Attribution-Share Alike 4.0 License. This regular paper is available in Fungal Genetics Reports: http://newprairiepress.org/fgr/vol52/iss1/4

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粗神经孢子虫最小庇护敲除突变体的构建

我一直在开发一种菌株，我认为这种菌株将有可能对基本基因进行常规的“最低限度的保护敲除”。它们同样可以被称为基本基因的“自我调节敲低”。这种最小保护的敲除可以在微阵列分析中得到比异核体或同源核体用可诱导的野生型等位基因保护的结果更少受人为影响的结果。这种方法解决了关于每个克隆基因的两个问题，这些基因从此被统称为“你最喜欢的基因”(yfg)。Yfg应该是作为耐潮霉素敲除基因而不能成功通过交叉的基因。(1) yfg是可有可无的基因吗?(2)如果发现yfg是必需的，那么yfgΔ敲除突变的“终端表型”是什么?我所说的“末端表型”是指介于yfgΔ和部分二倍体yfg+/Δ之间的异核体，其中部分二倍体是允许某种程度上不理想生长的最小比例。本作品采用知识共享署名-相同方式共享4.0许可协议。这篇常规论文可在真菌遗传学报告:http://newprairiepress.org/fgr/vol52/iss1/4

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Fungal Genetics Reports

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