Highly-efficient colony PCR method for red yeasts and its application to identify mutations within two leucine auxotroph mutants.
作者： Xinping Lin , Fan Yang , Yongjin Zhou , Zhiwei Zhu , Guojie Jin
DOI: 10.1002/YEA.2926
关键词:
摘要: Red yeasts hold great promise in the production of microbial lipids and carotenoids. Genetic study red has attracted much attention; however, rapid amplification genes from yeast samples remains technically challenging. Here a highly efficient method for preparation genomic DNA (gDNA) template, which could be directly used PCR, was developed. Cells colonies or liquid cultures were collected sequentially treated by microwave, plMAN5C, proteinase K boiling (MMPB) single tube to give cell lysates that qualified as PCR templates. Single-copied gDNA fragments o up 2.8 kb successfully amplified. We also demonstrated successful application this species Ascomycetes Basidiomycetes identification two leucine auxotroph mutants Rhodotorula glutinis. This widely employed screening genetic engineering various yeasts.
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