| RRC ID |
47624
|
| Author |
Lin X, Yang F, Zhou Y, Zhu Z, Jin G, Zhang S, Zhao ZK.
|
| Title |
Highly-efficient colony PCR method for red yeasts and its application to identify mutations within two leucine auxotroph mutants.
|
| Journal |
Yeast
|
| Abstract |
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.
|
| Volume |
29(11)
|
| Pages |
467-74
|
| Published |
2012-11-1
|
| DOI |
10.1002/yea.2926
|
| PMID |
23065821
|
| MeSH |
Ascomycota / genetics
Basidiomycota / genetics
Biosynthetic Pathways / genetics
DNA, Fungal / genetics*
DNA, Fungal / isolation & purification*
Endopeptidase K / metabolism
Genetics, Microbial / methods*
Leucine / metabolism
Microwaves
Molecular Biology / methods*
Mutation*
Polymerase Chain Reaction / methods*
Rhodotorula / genetics*
|
| IF |
3.143
|
| Times Cited |
5
|
|
WOS Category
|
MYCOLOGY
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
MICROBIOLOGY
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| General Microbes |
JCM 8242 |
