Zeocin resistance as a dominant selective marker for transformation and targeted gene deletions in Candida glabrata
Zeocin resistance as a dominant selective marker for transformation and targeted gene deletions in Candida glabrata
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DOI：
10.1111/j.1439-0507.2006.01271.x
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发表时间：
2006-11-01
期刊：
影响因子：
4.9
通讯作者：
Tuite, Mick F.
中科院分区：
文献类型：
--
作者：
Alderton, Alex J.;Burr, Ian;Tuite, Mick F.
许多用于在光滑念珠菌中产生基因敲除的遗传工具利用营养缺陷型标记，但这不适合用于临床菌株。然而，抗生素抗性标记允许人们在没有对临床分离株进行任何先前遗传操作的情况下将靶基因删除。已经广泛报道了这种抗生素选择标记用于酿酒酵母的操作。然而，很少有抗生素抗性标记已被证明是有用的，在C。光滑的在这里，我们报道了利用来自印度斯坦链霉菌的ble基因编码的Zeocin抗性（Zeo（R））作为一种新的正选择标记用于C.包括临床菌株在内的光滑菌株对Zeocin比对G418显著更敏感。Zeo（R）标记在C. glabrata通过在实验室和临床菌株中构建ADE 2的缺失来证实。glabrata，使用短（90 bp）和长（400 bp）同源盒。
Many of the genetic tools used to generate gene knockouts in Candida glabrata exploit auxotrophic markers but this is not suitable for use with clinical strains. Antibiotic resistance markers, however, allow one to target genes to be deleted without any prior genetic manipulation of clinical isolates. Such antibiotic selection markers have been widely reported for the manipulation of Saccharomyces cerevisiae. However, very few antibiotic resistance markers have been shown to be useful in C. glabrata. Here, we report the use of Zeocin resistance (Zeo(R)), encoded by the ble gene from Streptoalloteichus hindustanus, as a new positive selection marker for the genetic manipulation of C. glabrata including clinical strains that we show are significantly more sensitive to Zeocin than to G418. The potential of the Zeo(R) marker for targeted gene disruption in C. glabrata was confirmed by constructing deletions of the ADE2 in both a laboratory and a clinical strain of C. glabrata, using both short (90 bp) and long (400 bp) homology cassettes.