Hentges, P., Van Driessche, B., Tafforeau, L., Vandenhaute, J. and Carr, A. M. (2005) Three novel antibiotic marker cassettes for gene disruption and marker switching in Schizosaccharomyces pombe. Yeast, 22 (13). pp. 1013-9. ISSN 0749-503xFull text not available from this repository.
The ease of construction of multiple mutant strains in Schizosaccharomyces pombe is limited by the number of available genetic markers. We describe here three new cassettes for PCR-mediated gene disruption that can be used in combination with commonly used fission yeast markers to make multiple gene deletions. The natMX6, hphMX6 and bleMX6 markers give rise to resistance towards the antibiotics nourseothricin (NAT), hygromycin B and phleomycin, respectively. The cassettes are composed of exogenous sequences to increase the frequency of integration at targeted loci, and have a structure similar to the commonly used pFA6a-kanMX6 modular plasmid system. This allows a simple exchange of the kanMX6 marker in existing strains with any of the three new cassettes. Alternatively, oligonucleotide primers designed for the modular kanMX6 cassettes can be used to make the transforming PCR fragments for gene disruption. We illustrate the construction of a mutant strain with six independent gene disruptions, using the novel antibiotic cassettes in combination with existing genetic markers. Copyright (c) 2005 John Wiley & Sons, Ltd.
|Additional Information:||0749-503x Journal Article|
|Schools and Departments:||School of Life Sciences|
|Depositing User:||Gee Wheatley|
|Date Deposited:||19 Mar 2007|
|Last Modified:||30 Nov 2012 16:51|
|Google Scholar:||58 Citations|