Nonconventional yeast Candida famata and Ogataea polymorpha are interesting organisms for basic and applied studies. O. polymorpha is methylotrophic thermotolerant yeast capable of xylose alcoholic fermentation whereas C. famata is capable of riboflavin overproduction. Still, the new tools for molecular research of these species are needed. The aim of this study was to develop the new dominant selective markers for C. famata and O. polymorpha usable in metabolic engineering experiments. In this work, the BSD gene from Aspergillus terreus coding for blasticidin S deaminase, O. polymorpha AUR1 gene required for sphingolipid synthesis and IMH3 gene, which encodes IMP dehydrogenase, were tested as the new dominant selective marker genes. Our results showed that AUR1 and IMH3 genes could be used as dominant selective markers for O. polymorpha with frequencies of transformation of 40 and 20 transformants per microgram of DNA, respectively. The IMH3 gene was successfully used as the marker for construction of O. polymorpha strains with increased ethanol production from xylose due to overexpression of TAL1, TKL1 and AOX1 genes. The BSD gene from A. terreus, conferring resistance to blasticidin, was found to be efficient for selection of C. famata transformants.

中文翻译：

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非常规酵母Ogataea polymorpha和Candida famata的新的显性选择标记的开发

非常规的酵母假丝酵母和多形的Ogataea是有趣的生物，可用于基础和应用研究。O. polymorpha是能够进行木糖酒精发酵的甲基营养型耐高温酵母，而C. famata能够生产核黄素。尽管如此，仍需要用于这些物种的分子研究的新工具。这项研究的目的是开发新的显性选择标记C. famata和O.多形代谢工程实验使用。在这项工作中，来自曲霉曲霉的BSD基因编码稻瘟菌素S脱氨酶O.polymorpha AUR1鞘脂合成所需的基因和编码IMP脱氢酶的IMH3基因被测试为新的显性选择标记基因。我们的结果表明，AUR1和IMH3基因可以用作O. polymorpha的主要选择标记，每微克DNA的转化频率分别为40和20个转化子。所述IMH3基因成功用作标记用于建筑的O.多形与来自木糖的乙醇生产提高的菌株由于过表达TAL1，TKL1和AOX1基因。土曲霉的BSD基因赋予对杀稻瘟菌素的抗性，对于选择C.famata转化体是有效的。