Reconstitution of RNase P activity from inactive RNA and protein.
Author(s) -
Ryszard Kole,
Sidney Altman
Publication year - 1979
Publication title -
proceedings of the national academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.76.8.3795
Subject(s) - rnase p , rna , sephadex , urea , escherichia coli , biochemistry , degradosome , chemistry , rnase ph , nuclease protection assay , chromatography , biology , microbiology and biotechnology , enzyme , non coding rna , gene
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.
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