Characterization of DNA polymerase φ from Saccharomyces cerevisiae

酿酒酵母 DNA 聚合酶 φ 的表征

• 批准号: 15570144
• 负责人: SHIMIZU Kikuo
• 金额: $ 2.37万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15570144/
• 关键词: yeast; DNA microarray; RNA; transcription; たんぱく発現

The new type mutant (degron mutant) was constructed in order to analyze the function of DNA polymerase φ in vivo. The degron mutant of POL5 (pol5^<td>) has the peptide which is substrate of proteolysis system of the ubiquitin in the upstream of the gene of DNA polymerase φ. This mutant also has plasmid for large manifesting UBC4 gene product of the degrading system, because it did not become the lethality in the high temperature (37 degrees) without overproducing the UBC4 gene product.The DNA microarray analysis of the POL5 gene was carried out in order to examine the pattern of the gene expression at non-permissive temperature using the mutant pol5^<td> of DNA polymerase φ, and it was compared with the expression of the wild type by same condition. The result showed that the specific gene cluster peculiarly changed was not obtained.This fact means that POL5 gene carries out universal transcriptional control not but specific gene group.

为了分析DNA聚合酶φ的体内功能，构建了一种新型突变体（降解决定子突变体）。POL5的降解决定子突变体（pol5^<td>）在DNA聚合酶φ基因的上游具有泛素蛋白水解系统的底物肽。该突变体也具有用于大量显示降解系统的UBC 4基因产物的质粒，因为它在高温（37 ℃）下没有过量产生UBC 4基因产物而没有成为致死性。为了使用DNA聚合酶φ的突变体pol 5 ^检查在非允许温度下的基因表达模式，进行了POL 5基因的DNA微阵列分析<td>，并将其与相同条件下野生型的表达进行了比较。结果表明，POL5基因没有得到特异性变化的基因簇，这说明POL5基因进行的不是普遍的转录调控，而是特定的基因组。

项目成果

DNA polymerases from the budding yeast Saccharomyces cerevisiae : Prospect on fidelity and mutagenesis caused by DNA polymerases

来自芽殖酵母酿酒酵母的 DNA 聚合酶：DNA 聚合酶引起的保真度和诱变的前景

• 发表时间: 2004
• 期刊: Recent.Res.Devel.Biochem. 5
• 作者: Shimizu;Kikuo
• 通讯作者: Kikuo