Sasaki T
Fermentation Research Laboratories, Sankyo Co., Ltd., Tokyo, Japan.
Appl Environ Microbiol. 1992 Mar;58(3):948-52. doi: 10.1128/aem.58.3.948-952.1992.
Cells of an asporogenous industrial strain of the yeast Saccharomyces cerevisiae were irradiated with UV light by using a method that was developed previously (T. Sasaki and Y. Ohshima, Appl. Environ. Microbiol. 53:1504-1511, 1987). This treatment gave rise to large-cell clones among the surviving cells, from which colonies consisting of cells with a normal morphology and a prototrophic property were obtained. The large-cell trait of these was stably inheritable, with the cell volumes being about twice that of the parent for 7 years on a slant agar medium at 4 degrees C with repeated transfers. The cellular DNA content of these clones, in comparison to those of two authentic haploid strains, was determined by chemical analysis. The ratio of the DNA contents showed that the parent and its large-cell derivatives were a diploid and tetraploids, respectively. No abnormality was found in the chromosomal DNA patterns of the large-cell clones, at least as determined by agarose gel electrophoresis with a CHEF-DR II pulsed-field electrophoresis system. These findings led to the conclusion that our UV light method is applicable for inducing ploidy level increments in the widely used yeast species S. cerevisiae.
采用先前开发的方法(T. Sasaki和Y. Ohshima,《应用与环境微生物学》53:1504 - 1511,1987年),用紫外线照射酿酒酵母的一个不产孢子的工业菌株的细胞。这种处理在存活细胞中产生了大细胞克隆,从中获得了由具有正常形态和原养型特性的细胞组成的菌落。这些细胞的大细胞特性是稳定可遗传的,在4℃的斜面琼脂培养基上反复传代培养7年,其细胞体积约为亲本的两倍。通过化学分析确定了这些克隆的细胞DNA含量,并与两个正宗单倍体菌株的DNA含量进行了比较。DNA含量的比值表明,亲本及其大细胞衍生物分别为二倍体和四倍体。至少用CHEF - DR II脉冲场电泳系统进行琼脂糖凝胶电泳测定时,大细胞克隆的染色体DNA图谱未发现异常。这些发现得出结论,我们的紫外线方法适用于在广泛使用的酿酒酵母物种中诱导倍性水平增加。