A counterselection for the tryptophan pathway in yeast: 5‐fluoroanthranilic acid resistance

The ability to counterselect, as well as to select for, a genetic marker has numerous applications in microbial genetics. Described here is the use of 5‐fluoroanthranilic acid for the counterselection of TRP1, a commonly used genetic marker in the yeast Saccharomyces cerevisiae. Counterselection usi...

Full description

Saved in:
Bibliographic Details
Published in:Yeast (Chichester, England) England), 2000-04, Vol.16 (6), p.553-560
Main Authors: Toyn, Jeremy H., Gunyuzlu, Paul L., Hunter White, W., Thompson, Lorin A., Hollis, Gregory F.
Format: Article
Language:English
Subjects:
5-fluoroanthranilic acid
Aldose-Ketose Isomerases
Anthranilate Phosphoribosyltransferase - genetics
Anthranilate Synthase - genetics
antimetabolism
Cell Division - drug effects
Cell Division - genetics
counterselection
Drug Resistance, Microbial - genetics
fluoroanthranilic acid
Fungal Proteins - genetics
Gene Deletion
Genetic Markers
Indole-3-Glycerol-Phosphate Synthase - genetics
Mutation
ortho-Aminobenzoates - chemistry
ortho-Aminobenzoates - pharmacology
Plasmids - genetics
Recombinant Fusion Proteins - genetics
Saccharomyces cerevisiae
Saccharomyces cerevisiae - drug effects
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - metabolism
Saccharomyces cerevisiae Proteins
TRP1
TRP1 gene
tryptophan
Tryptophan - biosynthesis
Tryptophan Synthase - genetics
yeast
Citations: Items that this one cites
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The ability to counterselect, as well as to select for, a genetic marker has numerous applications in microbial genetics. Described here is the use of 5‐fluoroanthranilic acid for the counterselection of TRP1, a commonly used genetic marker in the yeast Saccharomyces cerevisiae. Counterselection using 5‐fluoroanthranilic acid involves antimetabolism by the enzymes of the tryptophan biosynthetic pathway, such that trp1, trp3, trp4 or trp5 strains, which lack enzymes required for the conversion of anthranilic acid to tryptophan, are resistant to 5‐fluoroanthranilic acid. Commonly used genetic procedures, such as selection for loss of a chromosomally integrated plasmid, and a replica‐plating method to rapidly assess genetic linkage in self‐replicating shuttle vectors, can now be carried out using the TRP1 marker gene. In addition, novel tryptophan auxotrophs can be selected using 5‐fluoroanthranilic acid. Copyright © 2000 John Wiley & Sons, Ltd.
ISSN:0749-503X
1097-0061
DOI:10.1002/(SICI)1097-0061(200004)16:6<553::AID-YEA554>3.0.CO;2-7