Cloning of Saccharomyces cerevisiae DNA replication genes: isolation of the CDC8 gene and two genes that compensate for the cdc8-1 mutation.
Cloning of Saccharomyces cerevisiae DNA replication genes: isolation of the CDC8 gene and two genes that compensate for the cdc8-1 mutation.
复制标题
酿酒酵母 DNA 复制基因的克隆：分离 CDC8 基因和两个补偿 cdc8-1 突变的基因。
DOI：
10.1128/mcb.3.10.1730-1737.1983
复制
发表时间：
1983
影响因子：
5.3
通讯作者：
Campbell,JL
中科院分区：
文献类型：
--
作者：
Kuo,CL;Campbell,JL
CDC8基因的产物是酿酒酵母DNA复制所必需的，已在重组质粒上分离出来。带有酵母ARS1、CEN4和URA3序列的酵母载体YCp50分别用于制备包含整个酵母基因组的重组质粒池，以提供复制、稳定性和选择。通过转化 acdc8-1 突变体并选择能够在非允许温度下生长的克隆，分离出能够补充温度敏感 cdc8-1 突变的质粒。删除图谱显示，整个互补活性是在 0.75 KB 的片段上进行的。该片段位于已充分表征的sup4基因下游1kb处，该基因已知与CDC8有遗传连锁，从而证实克隆的基因对应于染色体CDC8基因。本研究还鉴定了另外两个重组质粒，它们补充了 cdc8-1 突变，但不包含 0.75 KB 片段或任何侧翼 DNA。这些质粒可以含有补偿CDC8基因产物的缺乏的基因。
TheCDC8gene, whose product is required for DNA replication inSaccharomyces cerevisiae, has been isolated on recombinant plasmids. The yeast vector YCp50 bearing the yeastARS1,CEN4, andURA3sequences, to provide for replication, stability, and selection, respectively, was used to prepare a recombinant plasmid pool containing the entire yeast genome. Plasmids capable of complementing the temperature-sensitivecdc8-1mutation were isolated by transformation of acdc8-1mutant and selection for clones able to grow at the nonpermissive temperature. The entire complementing activity is carried on a 0.75-kilobase fragment, as revealed by deletion mapping. This fragment lies 1 kilobase downstream from the well-characterizedsup4gene, a gene known to be genetically linked toCDC8, thus confirming that the cloned gene corresponds to the chromosomalCDC8gene. Two additional recombinant plasmids that complement thecdc8-1mutation but that do not contain the 0.75-kilobase fragment or any flanking DNA were also identified in this study. These plasmids may contain genes that compensate for the lack ofCDC8gene product.