Zillig W, Stetter K O, Tobien M
Eur J Biochem. 1978 Nov 2;91(1):193-9. doi: 10.1111/j.1432-1033.1978.tb20951.x.
DNA-dependent RNA polymerase core enzyme was isolated from Halobacterium halobium. The purification is based on the finding that the enzyme is stable in 40% (v/v) glycerol, in the presence of 0.05 M MgCl2 and involves adsorption of contaminants to DEAE-cellulose, precipitation of the complex of polymerase with DNA by streptomycin sulfate, chromatography over Biogel and affinity chromatography over heparin-Sepharose or heparin-cellulose. The enzyme consists of four or five different subunits. The composition formula was estimated as (150000) (86000)2 (72000)2 (49000)3 or 2; there may be one or two different 49000-Mr subunits. RNA synthesis requires a template. Denatured DNA is more efficient than native DNA. The transcription of native DNA is specifically stimulated by the addition of a possibly sigma-like factor eluted from DEAE-cellulose. The fidelity of transcription is indicated by the absolute requirement for UTP besides ATP with poly[d(A-T)] as the template.
从嗜盐栖热菌中分离出了依赖DNA的RNA聚合酶核心酶。纯化过程基于以下发现：该酶在含有0.05 M MgCl2的40%（v/v）甘油中稳定，且涉及污染物吸附到DEAE - 纤维素上，用硫酸链霉素沉淀聚合酶与DNA的复合物，通过Biogel进行层析以及通过肝素 - 琼脂糖或肝素 - 纤维素进行亲和层析。该酶由四个或五个不同的亚基组成。其组成公式估计为(150000)(86000)2(72000)2(49000)3或2；可能存在一个或两个不同的49000 - Mr亚基。RNA合成需要模板。变性DNA比天然DNA更有效。通过添加从DEAE - 纤维素上洗脱的一种可能类似σ因子，可特异性刺激天然DNA的转录。以聚[d(A - T)]为模板时，除了ATP外对UTP的绝对需求表明了转录的保真度。
