NDLI: Differential activity staining: its use in characterization of guanylyl-specific ribonuclease in the genus Ustilago.

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Differential activity staining: its use in characterization of guanylyl-specific ribonuclease in the genus Ustilago.

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Differential activity staining: its use in characterization of guanylyl-specific ribonuclease in the genus Ustilago.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Blank, A. Dekker, C. A.
Abstract	Guanylyl-specific ribonuclease can be identified by a novel technique employing electrophoresis in polyacrylamide slabs followed by differential activity staining. The technique requires as little as 7 ng of enzyme which may be grossly admixed with contaminants, including other ribonucleases. Upon electrophoresis and activity staining, a variety of ribonucleases can be visualized as light or clear bands in a colored background formed by toluidine blue complexed with oligonucleotide substrate. Guanylyl-specific ribonuclease, which is detectable when using an oligonucleotide substrate of random base sequence, does not yield a band when using oligonucleotides bearing guanylyl residues at the 3'-termini only and containing, therefore, no susceptible internucleotide bonds; in contrast, a ribonuclease with a different base specificity or no base specificity yields a band with either substrate. This differential activity staining method for establishing guanylyl specificity permits estimation of the extent of nonspecific cleavage of internucleotide linkages by a putatively guanylyl-specific enzyme and is at least as sensitive as conventional procedures for determination of base specificity. With this new technique guanyloribonuclease has been identified in the unfractionated culture medium of 10 organisms belonging to the phytopathogenic fungal genus Ustilago. It is suggested that guanylyl-specific ribonuclease is widely distributed among Ustilago species; its electrophoretic properties may be revealing of phylogenetic relationships among these plant parasites and among their hosts. The general technique of differential activity staining, developed for determination of the base specificity of ribonucleases, may be widely applicable to analysis of enzymes catalyzing depolymerization reactions.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	12
Volume Number	72
Language	English
Publisher	National Academy of Sciences
Publisher Date	1975-01-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Basidiomycota Enzymology Ribonuclease T1 Metabolism Ribonucleases Ustilago Electrophoresis, Polyacrylamide Gel Indicators And Reagents Oligoribonucleotides Species Specificity Research Support, U.S. Gov't, Non-P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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