Cycloheximide-resicetance gene isolated from Candida maltosa : Mechanism of action and its utilization

麦芽糖念珠菌中分离的放线菌酮抗性基因：作用机制及其利用

• 批准号: 61470129
• 负责人: TAKAGI Masamichi
• 金额: $ 2.82万
• 依托单位: Faculty of Agriculture, The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1986
• 资助国家: 日本
• 起止时间: 1986 至 1987
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-61470129/
• 关键词: Candida maltosa; Saccharomyces cerevisiae; cycloheximide; gene cloning; protein syntesis; ribosome modification; Saccharomyses cerevisiae; リボソーム; 抗生物質耐性; 耐性化遺伝子; クローニング

1. We isolated a gene designated the RIM-C gene (ribosome-modification by cycloheximide) from the genome of Candida maltosa by using a host-vector system of Sachharomyces cerevisiae, which is sensitive to cycloheximide (CYH). When this gene was introduced to S. cerevisiae, its ribosomes were modified to be resistant to CYH.2. We determined the nucleotide sequence of DNA region containing the RIM-C gene.3. By using the dot-hybridization technique, we found that the modified ribosomes did not contain a RNA product of the gene. An RNA product of this gene was found to contain poly(A)-tail. These data indicate that the gene encode a mRNA and not a new species of rRNA.4. By using the two-dimensional gel electrophoresis, we did not found a new ribosomal protein in the modified ribosomes. It is suggested that the gene does not code for a new ribosomal protein, but codes for a modification enzymes of a component of ribosomes.5. We determined the transcription strand of the gene and the transcription start-point.

1.我们使用酿酒酵母的宿主-载体系统从麦芽糖假丝酵母基因组中分离出一个对放线菌酮敏感的RIM-C基因（核糖体修饰放线菌酮）。将该基因导入S. cerevisiae，其核糖体被修饰以抵抗CYH。确定了RIM-C基因所在DNA区域的核苷酸序列.通过使用斑点杂交技术，我们发现修饰的核糖体不包含该基因的RNA产物。发现该基因的RNA产物含有poly（A）尾。这些数据表明，该基因编码的是mRNA，而不是一种新的rRNA。双向凝胶电泳结果表明，修饰后的核糖体中没有发现新的核糖体蛋白。这表明该基因不是编码新的核糖体蛋白，而是编码核糖体组分的修饰酶。我们确定了该基因的转录链和转录起点。

项目成果

高木正道他: J. Gen. Appl. Microbiol. 31. 267-275 (1985)

M. Takagi 等人：《微生物学杂志》31. 267-275 (1985)

高木正道: J. Bacteriol. 168. 417-419 (1986)

高木正通：细菌学杂志 168. 417-419 (1986)

Masamichi,Takagi et al.: "Cloning in Saccharomyces cerevisiae of a cycloheximide resistance gene from the Candida maltosa which modifies ribosomes" J. Bacteriol.168. 417-419 (1986)

Masamichi，Takagi 等人：“在酿酒酵母中克隆来自麦芽糖假丝酵母的放线菌酮抗性基因，该基因修饰核糖体”J. Bacteriol.168。

高木正道: J. Bacteriol.

高木正道：细菌学杂志。

高木正道 他: J.Bacteriol.168. 471-419 (1986)

Masamichi Takagi 等：J.Bacteriol.168（1986）。