Kozhina T N, Peshekhonov V T, Chepurnaia O V, Korolev V G
Mol Gen Mikrobiol Virusol. 1991 Mar(3):13-6.
A new convenient system for isolation of the yeast mutants deficient in the genetical recombination is proposed. The chimeric plasmids constructed to carry the noncomplimenting mutant copy of the yeast ADE2 gene and different selectable yeast markers (LEU2 or TRP1 genes) are the basis for the system. Interplasmid intragenic recombination of ADE2 gene alleles in yeast cells transformed by two chimeric plasmids results in appearance of the secondary white prototrophic clones covering the primary red colony. The number of the clones reflects the recombination processes and is subject to an easy visual control. The proposed technique allows one to reveal both hypo and hyperrecombination mutants. Crossover or the gene conversion events can be distinguished by the simple genetical analysis of the secondary clones. The collection of mutants deficient in the genetical recombination has been obtained by the proposed technique.
提出了一种用于分离基因重组缺陷型酵母突变体的新型便捷系统。构建携带酵母ADE2基因非互补突变拷贝和不同可选择酵母标记(LEU2或TRP1基因)的嵌合质粒是该系统的基础。由两个嵌合质粒转化的酵母细胞中ADE2基因等位基因的质粒间基因内重组导致覆盖原始红色菌落的次级白色原养型克隆出现。克隆数量反映了重组过程,易于目视控制。所提出的技术能够揭示低重组和高重组突变体。通过对次级克隆进行简单的遗传分析可以区分交叉或基因转换事件。利用所提出的技术已获得了基因重组缺陷型突变体库。