Unk Ildiko, Daraba Andreea
The Institute of Genetics, Biological Research Center of The Hungarian Academy of Sciences, Szeged, Hungary.
Bio Protoc. 2014 Oct 20;4(20). doi: 10.21769/BioProtoc.1272.
There are several methods to measure the capacity of yeast cell to respond to environmental impacts on their genome by mutating it. One frequently used method involves the detection of forward mutations in the gene. The gene encodes for an arginine permease that is responsible for the uptake of arginine and it can also transport the toxic analog of arginine, canavanine (Whelan , 1979). When cells are grown on a media containing canavanine but lacking arginine, the cells die because of the uptake of the toxic canavanine. However, if a mutation in the gene inactivates the permease, that cell survives and forms a colony on the plate. The following protocol describes the measurement of UV-induced mutagenesis at the locus.
有几种方法可用于测量酵母细胞通过使其基因组发生突变来响应环境影响的能力。一种常用的方法涉及检测该基因中的正向突变。该基因编码一种精氨酸通透酶，负责精氨酸的摄取，它也能转运精氨酸的有毒类似物刀豆氨酸（惠兰，1979年）。当细胞在含有刀豆氨酸但缺乏精氨酸的培养基上生长时，细胞会因摄取有毒的刀豆氨酸而死亡。然而，如果该基因发生突变使通透酶失活，该细胞就能存活并在平板上形成菌落。以下方案描述了在该位点测量紫外线诱导的诱变作用。
