McCusker J H, Haber J E
Department of Biology, Brandeis University, Waltham, Massachusetts 02254.
Genetics. 1988 Jun;119(2):303-15. doi: 10.1093/genetics/119.2.303.
We describe the isolation and preliminary characterization of a set of pleiotropic mutations resistant to the minimum inhibitory concentration of cycloheximide and screened for ts (temperature-sensitive) growth. These mutations fall into 22 complementation groups of cycloheximide resistant ts lethal mutations (crl). None of the crl mutations appears to be allelic with previously isolated mutations. Fifteen of the CRL loci have been mapped. At the nonpermissive temperature (37 degrees), these mutants arrest late in the cell cycle after several cell divisions. Half of these mutants are also unable to grow at very low temperatures (5 degrees). Although mutants from all of the 22 complementation groups exhibit similar temperature-sensitive phenotypes, an extragenic suppressor of the ts lethality of crl3 does not relieve the ts lethality of most other crl mutants. A second suppressor mutation allows crl10, crl12, and crl14 to grow at 37 degrees but does not suppress the ts lethality of the remaining crl mutants. We also describe two new methods for the enrichment of auxotrophic mutations from a wild-type yeast strain.
我们描述了一组对放线菌酮最低抑菌浓度具有抗性并经温度敏感(ts)生长筛选的多效性突变体的分离及初步特性分析。这些突变分为22个放线菌酮抗性ts致死突变(crl)互补群。没有一个crl突变似乎与先前分离的突变等位。已对15个CRL位点进行了定位。在非允许温度(37℃)下,这些突变体在经过几次细胞分裂后在细胞周期后期停滞。其中一半突变体在极低温度(5℃)下也无法生长。尽管来自所有22个互补群的突变体表现出相似的温度敏感表型,但crl3的ts致死性的一个基因外抑制子并不能缓解大多数其他crl突变体的ts致死性。第二个抑制突变使crl10、crl12和crl14在37℃下生长,但不能抑制其余crl突变体的ts致死性。我们还描述了两种从野生型酵母菌株中富集营养缺陷型突变的新方法。