Journal
YEAST
Volume 29, Issue 11, Pages 467-474Publisher
WILEY-BLACKWELL
DOI: 10.1002/yea.2926
Keywords
red yeast; screening; colony PCR; leucine auxotroph
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Funding
- Natural Sciences Foundation of China [31000052]
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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.
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