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多头绒泡菌两种RNA聚合酶的纯化及性质

Purification and properties of two RNA polymerases from Physarum polycephalum.

作者信息

Burgess A B, Burgess R R

出版信息

Proc Natl Acad Sci U S A. 1974 Apr;71(4):1174-7. doi: 10.1073/pnas.71.4.1174.

DOI:10.1073/pnas.71.4.1174
PMID:4524629
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC388186/
Abstract

Two RNA polymerases have been purified from the slime mold Physarum polycephalum, one sensitive and one resistant to alpha-amanitin. Both enzymes are more active with denatured DNA than native DNA as a template and prefer Mn(++) rather than Mg(++) as a divalent cation. The alpha-amanitin-sensitive enzyme shows maximum activity at 0.15 M KCl, whereas the resistant enzyme is most active at very low ionic strength. Analysis of the resistant enzyme on polyacrylamide gels containing sodium dodecyl sulfate shows two subunits present in a 1:1 ratio with molecular weights of 205,000 and 125,000.

摘要

已从多头绒泡菌中纯化出两种RNA聚合酶,一种对α-鹅膏蕈碱敏感,另一种对其具有抗性。作为模板,这两种酶对变性DNA的活性均高于天然DNA,并且更倾向于以Mn(++)而非Mg(++)作为二价阳离子。α-鹅膏蕈碱敏感的酶在0.15M KCl浓度下显示出最大活性,而抗性酶在极低离子强度下活性最高。在含有十二烷基硫酸钠的聚丙烯酰胺凝胶上对抗性酶进行分析,结果显示存在两个亚基,其比例为1:1,分子量分别为205,000和125,000。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c9be/388186/1c3e508efbd4/pnas00057-0170-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c9be/388186/1c3e508efbd4/pnas00057-0170-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c9be/388186/1c3e508efbd4/pnas00057-0170-a.jpg

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本文引用的文献

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Protein measurement with the Folin phenol reagent.使用福林酚试剂进行蛋白质测定。
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Specific nucleolar and nucleoplasmic RNA polymerases.特定的核仁与核质RNA聚合酶。
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Isolation and DNA content of nuclei of Physarum polycephalum.多头绒泡菌细胞核的分离及DNA含量
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A new method for the large scale purification of Escherichia coli deoxyribonucleic acid-dependent ribonucleic acid polymerase.一种大规模纯化大肠杆菌脱氧核糖核酸依赖性核糖核酸聚合酶的新方法。
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DNA dependent-RNA polymerases from Physarum polycephalum.多头绒泡菌的DNA依赖性RNA聚合酶
FEBS Lett. 1973 Sep 1;35(1):41-4. doi: 10.1016/0014-5793(73)80572-1.
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J Biol Chem. 1973 Jun 10;248(11):3933-9.