大肠杆菌中依赖脱氧核糖核酸的核糖核酸聚合酶的研究。生长过程中该酶活性的变化。
Studies on deoxyribonucleic acid-dependent ribonucleic acid polymerase from Escherichia coli. Variations of the enzyme activity during growth.
作者信息
Abraham K A, Andersen K J, Rognes A
出版信息
Biochem J. 1972 Sep;129(2):291-9. doi: 10.1042/bj1290291.
Abstract
- RNA polymerase activity of Escherichia coli extracts prepared from cells in exponential and stationary phases of growth, when measured in the presence and absence of external template, showed significant qualitative differences. 2. In both extracts, polymerase activity was higher when assayed with external template, suggesting the presence of a pool of enzyme not bound to cellular DNA. 3. In the crude extract, the fraction of enzyme bound to cellular DNA is higher during the exponential phase of growth. 4. A method is described for the purification of enzyme molecules not tightly bound to cellular DNA from exponential- and stationary-phase cultures. 5. Purified enzyme preparations showed differences in template requirement and subunit composition. 6. On phosphocellulose chromatography of stationary-phase enzyme, a major portion of polymerase activity eluted from the column with 0.25m-KCl. In the case of exponential-phase enzyme, polymerase activity eluted from a phosphocellulose column mainly with 0.35m-KCl. 7. Enzyme assays done with excess of bacteriophage T(4) DNA showed a strong inhibition of stationary-phase enzyme by this template. The exponential-phase enzyme was only slightly inhibited by excess of bacteriophage T(4) DNA.
摘要
- 对处于生长指数期和稳定期的大肠杆菌细胞提取物的RNA聚合酶活性进行检测，无论有无外部模板存在，均显示出显著的质量差异。2. 在两种提取物中，使用外部模板检测时聚合酶活性更高，这表明存在一组未与细胞DNA结合的酶。3. 在粗提取物中，生长指数期细胞中与细胞DNA结合的酶比例更高。4. 描述了一种从生长指数期和稳定期培养物中纯化未紧密结合细胞DNA的酶分子的方法。5. 纯化的酶制剂在模板需求和亚基组成方面存在差异。6. 对稳定期酶进行磷酸纤维素层析时，大部分聚合酶活性在0.25m - KCl条件下从柱上洗脱。对于指数期酶，聚合酶活性主要在0.35m - KCl条件下从磷酸纤维素柱上洗脱。7. 用过量噬菌体T(4) DNA进行的酶活性检测显示，该模板对稳定期酶有强烈抑制作用。过量噬菌体T(4) DNA对指数期酶仅有轻微抑制作用。
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