Cloning by gene amplification of two loci conferring multiple drug resistance in Saccharomyces.
Cloning by gene amplification of two loci conferring multiple drug resistance in Saccharomyces.
复制标题
通过基因扩增克隆酵母菌中赋予多重耐药性的两个基因座。
DOI：
10.1093/genetics/125.1.13
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发表时间：
1990
期刊：
影响因子：
3.3
通讯作者：
Golin,J
中科院分区：
文献类型：
--
作者：
Leppert,G;McDevitt,R;Falco,SC;VanDyk,TK;Ficke,MB;Golin,J
分离出扩增后赋予多重耐药性的酵母 DNA 片段。将含有克隆在高拷贝自主复制载体 YEp24 中的酵母基因组文库的细胞铺在含有放线菌酮的培养基上。由于质粒携带的抗性决定子，100 个放线菌酮抗性菌落中有 5 个对不相关的抑制剂甲磺隆具有交叉抗性。从这些抗性克隆中分离出的质粒在酵母基因组中包含两个不重叠的区域，现在称为 PDR4 和 PDR5（多效性耐药）。 PDR4 被定位到 XIII 号染色体，距离 LYS7 31.5 cM，距离着丝粒 9 cM。 PDR4 被定位到 ADE2 和 H1S3 之间的 XV 染色体。遗传分析表明，至少三个介导抑制剂耐药性的紧密连锁基因（PDR5、PDR2 和 SMR3）位于该区域。 PDR4 或 PDR5 基因中的插入突变并不致命，但 PDR5 中的插入会导致药物过敏表型。
Yeast DNA fragments that confer multiple drug resistance when amplified were isolated. Cells containing a yeast genomic library cloned in the high copy autonomously replicating vector, YEp24, were plated on medium containing cycloheximide. Five out of 100 cycloheximide-resistant colonies were cross-resistant to the unrelated inhibitor, sulfometuron methyl, due to a plasmid-borne resistance determinant. The plasmids isolated from these resistant clones contained two nonoverlapping regions in the yeast genome now designated PDR4 and PDR5 (for pleiotropic drug resistant). PDR4 was mapped to chromosome XIII, 31.5 cM from LYS7 and 9 cM from the centromere. PDR4 was mapped to chromosome XV between ADE2 and H1S3. Genetic analysis demonstrated that at least three tightly linked genes (PDR5, PDR2 and SMR3) that mediate resistance to inhibitors are located in this region. Insertion mutations in the either PDR4 or PDR5 genes are not lethal, but the insertion in PDR5 results in a drug-hypersensitive phenotype.