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首页> 外文期刊>Biochemistry >Purification and initial characterization of RNA polymerase from Thermus thermophilus strain HB8.
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Purification and initial characterization of RNA polymerase from Thermus thermophilus strain HB8.

机译：嗜热栖热菌HB8的RNA聚合酶的纯化和初步表征。

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Utilizing a novel and rapid two-column purification procedure, the DNA-dependent RNA polymerase (RNAP) from the thermophile, Thermus thermophilus HB8, was purified to electrophoretic homogeneity with a recovery of 65% (as determined by RNAP activity) in less than 2 days. The purified enzyme was characterized using DNA containing the lambdaP(R) promoter. KMnO(4) footprinting, abortive initiation assays, and the formation of the specific stalled elongation complex provide compelling evidence that T. thermophilus RNA polymerase can bind to DNA containing the lambdaP(R) promoter, form an open complex, and initiate transcription in a temperature-dependent manner. This evidence suggests that T. thermophilus RNAP possesses less intrinsic binding energy than E. coli RNAP. Instead, T. thermophilus relies on the high temperatures of its environment to provide the thermal energy required to stimulate open promoter complex formation, initiate transcription, and facilitate the conformational changes in RNA polymerase that result in nucleotide incorporation.
机译：利用新颖，快速的两柱纯化程序，将嗜热菌Thermus thermophilus HB8的DNA依赖性RNA聚合酶（RNAP）纯化至电泳均一，回收率在不到2％的程度上达到了65％（由RNAP活性确定）。天。使用含有启动子的DNA表征纯化的酶。 KMnO（4）足迹，流产起始测定和特定的停滞伸长复合物的形成提供了令人信服的证据，即嗜热性粒细胞RNA聚合酶可以与包含lambdaP（R）启动子的DNA结合，形成开放复合物，并在a中启动转录。温度依赖的方式。该证据表明，嗜热链球菌RNAP比大肠杆菌RNAP具有更少的固有结合能。相反，嗜热链球菌依靠其环境的高温来提供刺激开放的启动子复合物形成，启动转录并促进导致核苷酸掺入的RNA聚合酶构象变化所需的热能。

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