Journal article
Puromycin- and methotrexate-resistance cassettes and optimized Cre-recombinase expression plasmids for use in yeast
Yeast (Chichester, England), Vol.32(5), pp.423-438
05/01/2015
DOI: 10.1002/yea.3069
PMCID: PMC4454448
PMID: 25688547
Abstract
Here we expand the set of tools for genetically manipulating Saccharomyces cerevisiae. We show that puromycin-resistance can be achieved in yeast through expression of a bacterial puromycin-resistance gene optimized to the yeast codon bias, which in turn serves as an easy-to-use dominant genetic marker suitable for gene disruption. We have constructed a similar DNA cassette expressing yeast codon-optimized mutant human dihydrofolate reductase (DHFR), which confers resistance to methotrexate and can also be used as a dominant selectable marker. Both of these drug-resistant marker cassettes are flanked by loxP sites, allowing for their excision from the genome following expression of Cre-recombinase. Finally, we have created a series of plasmids for low-level constitutive expression of Cre-recombinase in yeast that allows for efficient excision of loxP-flanked markers. Copyright (c) 2015 John Wiley & Sons, Ltd.
Details
- Title: Subtitle
- Puromycin- and methotrexate-resistance cassettes and optimized Cre-recombinase expression plasmids for use in yeast
- Creators
- Chris MacDonald - University of IowaRobert C. Piper - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Yeast (Chichester, England), Vol.32(5), pp.423-438
- DOI
- 10.1002/yea.3069
- PMID
- 25688547
- PMCID
- PMC4454448
- NLM abbreviation
- Yeast
- ISSN
- 0749-503X
- eISSN
- 1097-0061
- Publisher
- Wiley
- Number of pages
- 16
- Grant note
- R01GM094830 / NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of General Medical Sciences (NIGMS) NIHRO1 GM058202 / National Institutes of Health (NIH); United States Department of Health & Human Services; National Institutes of Health (NIH) - USA 13POST 14710040 / American Heart Association (AHA); American Heart Association
- Language
- English
- Date published
- 05/01/2015
- Academic Unit
- Molecular Physiology and Biophysics; Medicine Administration; Internal Medicine
- Record Identifier
- 9984297495602771
Metrics
11 Record Views