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Widespread occurrence of chromosomal aneuploidy following the routine production of Candida albicans mutants

机译:常规生产白色念珠菌突变体后染色体非整倍性的广泛发生

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It has come to our attention that approximately 35% of >100 published microarray datasets, where transcript levels were compared between two different strains, exhibit some form of chromosome-specific bias. While some of these arose from the use of strains whose aneuploidies were not known at the time, in a worrisome number of cases the recombinant strains have acquired additional aneuploidies that were not initially present in the parental strain. The aneuploidies often affected a different chromosome than the one harboring the insertion site. The affected strains originated from either CAI-4, RM1000, BWP17 or SN95 and were produced through a variety of strategies. These observations suggest that aneuploidies frequently occur during the production of recombinant strains and have an effect on global transcript profiles outside of the afflicted chromosome(s), thus raising the possibility of unintended phenotypic consequences. Thus, we propose that all Candida albicans mutants and strains should be tested for aneuploidy before being used in further studies. To this end, we describe a new rapid testing method, based on a multiplex quantitative PCR assay, that produces eight bands of distinct sizes from either the left or right arms of each C. albicans chromosome.
机译:引起我们注意的是,在> 100个已发布的微阵列数据集中,大约有35%的基因在两种不同菌株之间进行了转录水平比较,并表现出某种形式的染色体特异性偏倚。尽管其中一些是由于使用当时不知道非整倍性的菌株而引起的,但在令人担忧的情况下,重组菌株还获得了最初不在亲本菌株中存在的其他非整倍性。非整倍体通常影响与具有插入位点的染色体不同的染色体。受影响的菌株源自CAI-4,RM1000,BWP17或SN95,并通过多种策略产生。这些观察结果表明,非整倍性经常在重组菌株的产生过程中发生,并且对患病染色体外部的总体转录物谱有影响,从而增加了意外表型后果的可能性。因此,我们建议所有白念珠菌突变体和菌株在用于进一步研究之前都应进行非整倍性测试。为此,我们描述了一种基于多重定量PCR分析的新的快速测试方法,该方法从每个白色念珠菌染色体的左臂或右臂产生八个大小不同的条带。

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