Molecular Cell Biology, Groningen Biomolecular Sciences and Biotechnology Institute, Kluyver Centre for Genomics of Industrial Fermentation, University of Groningen, Groningen, The Netherlands.
FEMS Yeast Res. 2012 May;12(3):271-8. doi: 10.1111/j.1567-1364.2011.00772.x. Epub 2011 Dec 23.
Hansenula polymorpha is an important yeast in industrial biotechnology. In addition, it is extensively used in fundamental research devoted to unravel the principles of peroxisome biology and nitrate assimilation. Here we present an overview of key components of the genetic toolbox for H. polymorpha. In addition, we present new selection markers that we recently implemented in H. polymorpha. We describe novel strategies for the efficient creation of targeted gene deletions and integrations in H. polymorpha. For this, we generated a yku80 mutant, deficient in non-homologous end joining, resulting in strongly enhanced efficiency of gene targeting relative to the parental strain. Finally, we show the implementation of Gateway technology and a single-step PCR strategy to create deletions in H. polymorpha.
多形汉逊酵母是工业生物技术中的一种重要酵母。此外，它还广泛用于基础研究，以揭示过氧化物酶体生物学和硝酸盐同化的原理。本文概述了多形汉逊酵母遗传工具盒的关键组成部分。此外，我们还介绍了我们最近在多形汉逊酵母中实现的新选择标记。我们描述了在多形汉逊酵母中高效创建靶向基因缺失和整合的新策略。为此，我们生成了一个 yku80 突变体，该突变体缺乏非同源末端连接，从而相对于亲本菌株显著提高了基因靶向的效率。最后，我们展示了 Gateway 技术和单步 PCR 策略在多形汉逊酵母中创建缺失的应用。
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