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由无活性的RNA和蛋白质重建核糖核酸酶P活性。

Reconstitution of RNase P activity from inactive RNA and protein.

作者信息

Kole R, Altman S

出版信息

Proc Natl Acad Sci U S A. 1979 Aug;76(8):3795-9. doi: 10.1073/pnas.76.8.3795.

Abstract

RNase P preparations from Escherichia coli can be separated into RNA and protein by chromatography, in buffers containing 7 M urea, on Sephadex G-200, DEAE-Sephadex, or CM-Sephadex columns. Neither RNA nor protein components alone exhibits any RNase activity. RNase P activity can be reconstituted by mixing separated RNA and protein components in buffer containing 7M urea followed by dialysis of this mixture to remove the urea. Of several purified RNAs tried, only M2 RNA, the RNA species found in purified RNase P, is active in the reconstitution experiments.

摘要

通过在含有7M尿素的缓冲液中,在葡聚糖凝胶G - 200、二乙氨基乙基葡聚糖凝胶(DEAE - Sephadex)或羧甲基葡聚糖凝胶(CM - Sephadex)柱上进行色谱分离,可将来自大肠杆菌的核糖核酸酶P制剂分离为RNA和蛋白质。单独的RNA和蛋白质组分均不表现出任何核糖核酸酶活性。通过在含有7M尿素的缓冲液中混合分离的RNA和蛋白质组分,然后对该混合物进行透析以除去尿素,可重建核糖核酸酶P活性。在尝试的几种纯化RNA中,只有M2 RNA(在纯化的核糖核酸酶P中发现的RNA种类)在重建实验中具有活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d97e/383921/5ce1060d784d/pnas00008-0232-a.jpg

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