Transformation of protoplasted yeast cells is directly associated with cell fusion
Transformation of protoplasted yeast cells is directly associated with cell fusion
复制标题
原生质体酵母细胞的转化与细胞融合直接相关
DOI：
10.1128/mcb.4.4.771-778.1984
复制
发表时间：
1984
影响因子：
5.3
通讯作者：
Y. Oshima
中科院分区：
文献类型：
--
作者：
S. Harashima;A. Takagi;Y. Oshima
用两种方法估计了用带有染色体复制子（YRp或YCp）或2 μ DNA（YEp）的各种酵母质粒转化酵母原生质体期间的细胞融合频率。在一种方法中，转化了具有相同交配型和互补营养缺陷型核标记（有或没有细胞质标记）的两个单倍体菌株的原生质体混合物。当用从二项式分布理论导出的方程分析核标记的转化体的各种表型类别的数目时，在用YRp或YCp质粒的转化中，转化体之间的核融合频率为42 - 100%，而用YEp质粒的转化中为28 - 39%。在另一种方法中，用质粒转化携带sir突变的单倍体，sir突变允许MAT（交配型）基因座纯合的二倍体（或多倍体）通过沉默交配型基因座HML和HMR的表达形成孢子。在43%至95%的具有YRp或YCp质粒的转化体和26%至31%的YEp转化体中发现孢子形成能力。当细胞质混合包括核融合时，发现96%至100%的转化体是细胞融合体。基于这些观察，我们得出结论，酵母原生质体的转化与细胞融合直接相关。
The frequency of cell fusion during transformation of yeast protoplasts with various yeast plasmids with a chromosome replicon (YRp or YCp) or 2 mu DNA (YEp) was estimated by two methods. In one method, a mixture of protoplasts of two haploid strains with identical mating type and complementary auxotrophic nuclear markers with or without cytoplasmic markers was transformed. When the number of various phenotypic classes of transformants for the nuclear markers was analyzed by equations derived from binominal distribution theory, the frequency of nuclear fusion among the transformants was 42 to 100% in transformations with the YRp or YCp plasmids and 28 to 39% with the YEp plasmids. In another method, a haploid bearing the sir mutation, which allows a diploid (or polyploid) homozygous for the MAT (mating type) locus to sporulate by the expression of the silent mating-type loci HML and HMR, was transformed with the plasmids. Sporulation ability was found in 43 to 95% of the transformants with the YRp or YCp plasmids, and 26 to 31% of the YEp transformants. When cytoplasmic mixing was included with the nuclear fusion, 96 to 100% of the transformants were found to be cell fusants. Based upon these observations, we concluded that transformation of yeast protoplasts is directly associated with cell fusion.