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[特定寡核糖核苷酸与大肠杆菌RNA聚合酶结合的条件]

[Conditions for specific oligoribonucleotide binding with E. coli RNA-polymerase].

作者信息

Efimova L I, Knorre V L, Vasilenko S K, Savinkova L K, Salganik R I

出版信息

Mol Biol (Mosk). 1976 Mar-Apr;10(2):378-85.

PMID:781522
Abstract

RNA polymerase of E. coli (EC 2.7.7.6) is able to bind certain oligoribonucleotides with the length greater than or equal to 5 from the corresponding isoplith mixtures (Knorre V.L., Vasilenko S.V., Salganik R.I., FEBS Letters 30, 229, 1973). It has been shown in this study that all pentaribonucleotides able to be bound by RNA polymerase can be extracted from the random mixture by the enzyme saturation procedure. Loosely and tightly bound pentaribonucleotides subfractions were isolated and each was separated by chromatography into 3-4 isopliths. Blocking of the enzyme SH groups by p-chloromercurium benzoate (10(-3) M) and denaturation by urea (6.3 M) prevent formation of the enzyme-pentaribonucleotides complexes. Complexes are destroyed by heat denaturation. Removal of sigma-subunit does not influence the enzyme capacity for pentaribonucleotides binding.

摘要

大肠杆菌(EC 2.7.7.6)的RNA聚合酶能够与来自相应等长片段混合物中长度大于或等于5的某些寡核糖核苷酸结合(克诺尔V.L.、瓦西连科S.V.、萨尔加尼克R.I.,《欧洲生物化学学会联合会快报》30,229,1973)。本研究表明,所有能够被RNA聚合酶结合的五核糖核苷酸都可以通过酶饱和程序从随机混合物中提取出来。分离出了松散结合和紧密结合的五核糖核苷酸亚组分,并且每个亚组分都通过色谱法分离成3 - 4个等长片段。用对氯汞苯甲酸(10⁻³ M)封闭酶的SH基团以及用尿素(6.3 M)变性会阻止酶 - 五核糖核苷酸复合物的形成。复合物会因热变性而被破坏。去除σ亚基不会影响酶与五核糖核苷酸结合的能力。

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