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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.

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

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