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学科主题: 物理化学
题名: Highly-efficient colony PCR method for red yeasts and its application to identify mutations within two leucine auxotroph mutants
作者: Lin, Xinping1, 3;  Yang, Fan1, 2;  Zhou, Yongjin1, 3;  Zhu, Zhiwei1, 3;  Jin, Guojie1, 3;  Zhang, Sufang1;  Zhao, Zongbao Kent1
通讯作者: 张素芳
关键词: red yeast ;  screening ;  colony PCR ;  leucine auxotroph
刊名: YEAST
发表日期: 2012-11-01
DOI: 10.1002/yea.2926
卷: 29, 期:11, 页:467-474
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1816
产权排名: 1,1
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine
类目[WOS]: Biochemistry & Molecular Biology ;  Biotechnology & Applied Microbiology ;  Microbiology ;  Mycology
研究领域[WOS]: Biochemistry & Molecular Biology ;  Biotechnology & Applied Microbiology ;  Microbiology ;  Mycology
英文摘要: Red yeasts hold great promise in the production of microbial lipids and carotenoids. Genetic study of red yeasts has attracted much attention; however, rapid amplification of genes from red yeast samples remains technically challenging. Here a highly efficient method for the preparation of genomic DNA (gDNA) template, which could be directly used for PCR, was developed. Cells from colonies or liquid cultures were collected and sequentially treated by microwave, plMAN5C, proteinase K and boiling (MMPB) in a single tube to give cell lysates that were qualified as PCR templates. Single-copied gDNA fragments o up to 2.8?kb were successfully amplified. We also demonstrated successful application of this method for species in the Ascomycetes and Basidiomycetes and identification of two leucine auxotroph mutants of Rhodotorula glutinis. This method could be widely employed for the screening and genetic engineering of various yeasts. Copyright (c) 2012 John Wiley & Sons, Ltd.
关键词[WOS]: WALL LYTIC ENZYME ;  RHODOSPORIDIUM-TORULOIDES ;  CELL-WALL ;  SACCHAROMYCES-CEREVISIAE ;  IDENTIFICATION ;  PURIFICATION ;  DNA ;  EXTRACTION ;  CULTURE ;  LIPIDS
语种: 英语
WOS记录号: WOS:000310984900003
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/117863
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Dalian 116023, Peoples R China
2.Dalian Polytech Univ, Dalian, Peoples R China
3.Chinese Acad Sci, Grad Univ, Beijing, Peoples R China

Recommended Citation:
Lin, Xinping,Yang, Fan,Zhou, Yongjin,et al. Highly-efficient colony PCR method for red yeasts and its application to identify mutations within two leucine auxotroph mutants[J]. YEAST,2012,29(11):467-474.
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