Toyn J H, Gunyuzlu P L, White W H, Thompson L A, Hollis G F
Department of Applied Biotechnology, DuPont Pharmaceuticals Co., Wilmington, DE 19880, USA.
Yeast. 2000 Apr;16(6):553-60. doi: 10.1002/(SICI)1097-0061(200004)16:6<553::AID-YEA554>3.0.CO;2-7.
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.
对遗传标记进行反选择以及选择的能力在微生物遗传学中有众多应用。本文描述了使用5-氟邻氨基苯甲酸对TRP1进行反选择，TRP1是酿酒酵母中常用的遗传标记。使用5-氟邻氨基苯甲酸进行反选择涉及色氨酸生物合成途径的酶的抗代谢作用，使得缺乏将邻氨基苯甲酸转化为色氨酸所需酶的trp1、trp3、trp4或trp5菌株对5-氟邻氨基苯甲酸具有抗性。现在可以使用TRP1标记基因进行常用的遗传操作，例如选择染色体整合质粒的丢失，以及使用复制品平板法快速评估自我复制穿梭载体中的遗传连锁。此外，可以使用5-氟邻氨基苯甲酸选择新型色氨酸营养缺陷型。
