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Tools and methods for genetic analysis of Saccharomyces castellii

Astromskas, Eimantas LU and Cohn, Marita LU (2007) In Yeast 24(6). p.499-509
Abstract
The budding yeast species Saccharomyces castellii has provided important new insights into molecular evolution when incorporated in comparative genomics studies and studies of mitochondrial inheritage. Although it shows some diversity in the specific molecular details, several analyses have shown that it contains many genetic pathways similar to those of S. cerevisiae. Here we have investigated the possibility of performing genetic analyses in S. castellii. We optimized the LiAc transformation protocol to achieve 200-300 transformants/mu g plasmid DNA. We found that the commonly used plasmids for S. cerevisiae are stably maintained in S. castellii under selective conditions. Surprisingly, both 2 mu and CEN/ARS plasmids are kept at a high... (More)
The budding yeast species Saccharomyces castellii has provided important new insights into molecular evolution when incorporated in comparative genomics studies and studies of mitochondrial inheritage. Although it shows some diversity in the specific molecular details, several analyses have shown that it contains many genetic pathways similar to those of S. cerevisiae. Here we have investigated the possibility of performing genetic analyses in S. castellii. We optimized the LiAc transformation protocol to achieve 200-300 transformants/mu g plasmid DNA. We found that the commonly used plasmids for S. cerevisiae are stably maintained in S. castellii under selective conditions. Surprisingly, both 2 mu and CEN/ARS plasmids are kept at a high copy number. Moreover, the kanMX cassette can be used as a resistance marker against the selective drug geneticin (G418). Finally, we determined that the S. cerevisiae GAL] promoter can be used for the activation of transcription in S. castellii, thus enabling the controlled overexpression of genes when galactose is present in the medium. The availability of these tools provides the possibility or performing genetic analyses in S. castellii, and makes it a promising new model system in which hypotheses derived from bioinformatics studies can be experimentally tested. Copyright (C) 2007 John Wiley & Sons, Ltd. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
yeast, Saccharomyces, Saccharomyces castellii, Naumovia castellii, genetic tools, hemiascomycetes
in
Yeast
volume
24
issue
6
pages
499 - 509
publisher
John Wiley & Sons
external identifiers
  • wos:000247759800004
  • scopus:34347383977
ISSN
1097-0061
DOI
10.1002/yea.1488
language
English
LU publication?
yes
id
91a516a2-c3e2-4c8a-bca6-9f674c1d17e9 (old id 646071)
date added to LUP
2007-12-06 16:59:15
date last changed
2017-03-19 04:12:04
@article{91a516a2-c3e2-4c8a-bca6-9f674c1d17e9,
  abstract     = {The budding yeast species Saccharomyces castellii has provided important new insights into molecular evolution when incorporated in comparative genomics studies and studies of mitochondrial inheritage. Although it shows some diversity in the specific molecular details, several analyses have shown that it contains many genetic pathways similar to those of S. cerevisiae. Here we have investigated the possibility of performing genetic analyses in S. castellii. We optimized the LiAc transformation protocol to achieve 200-300 transformants/mu g plasmid DNA. We found that the commonly used plasmids for S. cerevisiae are stably maintained in S. castellii under selective conditions. Surprisingly, both 2 mu and CEN/ARS plasmids are kept at a high copy number. Moreover, the kanMX cassette can be used as a resistance marker against the selective drug geneticin (G418). Finally, we determined that the S. cerevisiae GAL] promoter can be used for the activation of transcription in S. castellii, thus enabling the controlled overexpression of genes when galactose is present in the medium. The availability of these tools provides the possibility or performing genetic analyses in S. castellii, and makes it a promising new model system in which hypotheses derived from bioinformatics studies can be experimentally tested. Copyright (C) 2007 John Wiley & Sons, Ltd.},
  author       = {Astromskas, Eimantas and Cohn, Marita},
  issn         = {1097-0061},
  keyword      = {yeast,Saccharomyces,Saccharomyces castellii,Naumovia castellii,genetic tools,hemiascomycetes},
  language     = {eng},
  number       = {6},
  pages        = {499--509},
  publisher    = {John Wiley & Sons},
  series       = {Yeast},
  title        = {Tools and methods for genetic analysis of Saccharomyces castellii},
  url          = {http://dx.doi.org/10.1002/yea.1488},
  volume       = {24},
  year         = {2007},
}