Purification and Characterization of RNA polymerase from halophilic archaebacteria.

嗜盐古细菌 RNA 聚合酶的纯化和表征。

• 批准号: 04808043
• 负责人: NAKAYAMA Masashi
• 金额: $ 1.02万
• 依托单位: Osaka University of Education
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04808043/
• 关键词: Halobacterium halobium; RNA polymerase; halophilic archaebacteria; Halobacterium halobium; RNA polymerase; halophilic archaebacteria

Halobacterium halobium, one of the halophilic archaebacteria, grows in extreme halophilic environments. The intracellular concentration of K^+ is also extremely high. The mechanisum of gene transcription in H.halobium and the nature of RNA polymerase involved in this process are still remained to study. We tried to purify the RNA polymerase which requires high salt concentrations for function.Zilling et al. (1978) identified a high-molecular-weight RNA polymerase from a related H.cutirubrum.In crude extract of H.halobium cells, we detected two RNA polymerase activities, one requiring high concentrations of salt for maximun activity and the other not requiring it. The high-requiring RNA polymerase was purified up to about 1,000-fold by PEI iteatment, Buty-Toyopearl and glycerol gradient centrifugation. The partially purified RNA polymerase required 3M NaCl for maximum activity DNA-dependent RNA synthesis continued at least until 90 min at 40ﾟC.The enzyme was resistant to both rifampicin and alpha-amanitin, but was sensitive to lignin, which is a potent inhibitor of influenza virus RNA polymerase. The molecular weight was estimated to be about 45 kDs by glycerol gradient centrifugation. Futher purification of the enzyme and detailed analysis of the enzyme structure are in progress.

卤素古细菌之一的盐酸盐杆菌在极端的卤素环境中生长。 K^+的细胞内浓度也非常高。 h钙中基因转录的机制和参与此过程的RNA聚合酶的性质仍然存在研究。我们试图纯化需要高盐浓度以进行功能的RNA聚合酶。 （1978年）从相关的H. cutirubrum.in中鉴定出高分子重量RNA聚合酶。通过PEI Iteatment，Buty-toyopearl和甘油梯度离心，将高压RNA聚合酶纯化至约1,000倍。部分纯化的RNA聚合酶需要3M NaCl才能使DNA依赖性RNA合成至少至少持续到40℃的90分钟C.该酶对Rifampicin和Alpha-Amanitin均具有抗性，但对木质素敏感，但对木质素敏感，这是造影症病毒RNA Polymase的潜在抑制剂。通过甘油梯度离心，估计分子量约为45 kds。酶的纯化和酶结构的详细分析正在进行中。