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Isolation of a haploid from an industrial Chinese rice wine yeast for metabolic engineering manipulation

机译：从工业黄酒酵母中分离单倍体以进行代谢工程操作

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摘要

The complex metabolic processes of yeast influence wine fermentation and therefore the quality of wine. Wine yeasts, owing to their being typically prototrophic and often polyploid, have been restricted in terms of exploiting classical recombinant genetic techniques to improve their characteristics. To overcome this problem, haploids have been isolated from a commercial Chinese rice wine strain N85, by disruption of the HO gene. In this study, the Cre-loxP system and a removable G418~r marker were used to construct an HO disruption cassette. Most of the heterologous sequences of constructed disruption cassette were successfully excised from the genome of the haploids by loop-out of the KanMX gene, through induced expression of the Cre recombinase. The removal of the resistant marker ensures the biological safety of the strains. As expected, no difference in fermentation capacity between the parental and the haploid strains was seen. The present work reports the construction of an HO disruption cassette by touchdown polymerase chain reaction and its application with a Chinese rice wine yeast for haploid isolation and to broaden physiological investigations and industrial applications.
机译：酵母的复杂代谢过程会影响葡萄酒的发酵，进而影响葡萄酒的质量。由于葡萄酒酵母通常是原养性的且通常是多倍体的，因此在利用经典重组遗传技术改善其特性方面受到了限制。为了克服这个问题，已经通过破坏HO基因从商业的中国黄酒菌株N85中分离了单倍体。在这项研究中，使用Cre-loxP系统和可移动的G418r标记构建HO破坏盒。通过Cre重组酶的诱导表达，通过环化KanMX基因，成功地从单倍体的基因组中切除了构建的破坏盒的大多数异源序列。去除抗性标记可确保菌株的生物学安全性。如预期的那样，亲本和单倍体菌株之间的发酵能力没有差异。目前的工作报道了通过触地聚合酶链反应构建HO破坏盒及其在中国黄酒酵母中用于单倍体分离的应用，并扩大了生理学研究和工业应用。

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  • 来源
    《Journal of the Institute of Brewing》 |2013年第4期|288-293|共6页
  • 作者

    Dianhui Wu; Xiaomin Li; Chao Shen; Jian Lu; Jian Chen; Guangfa Xie;

  • 作者单位

    The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi 214122, People's Republic of China,National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, People's Republic of China,School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, People's Republic of China;

    The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi 214122, People's Republic of China,National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, People's Republic of China,School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, People's Republic of China;

    The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi 214122, People's Republic of China,National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, People's Republic of China,School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, People's Republic of China;

    The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi 214122, People's Republic of China,National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, People's Republic of China,School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, People's Republic of China,Industrial Technology Research Institute of Jiangnan University in Suqian, 888 Renmin Road, Suqian 223800, People's Republic of China;

    The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi 214122, People's Republic of China,National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, People's Republic of China,School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, People's Republic of China;

    School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, People's Republic of China,National Engineering Research Centre for Chinese Rice Wine, China Shaoxing Rice Wine Group Co. Ltd, Shaoxing 312000, People's Republic of China;

  • 收录信息 美国《科学引文索引》(SCI);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    Chinese rice wine yeast; haploid; touchdown PCR; HO gene; Cre-loxP system;

    机译：中国黄酒酵母;单倍体;触地PCR;HO基因;Cre-loxP系统;

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