Affinity electrophoresis for monitoring terminal phosphorylation and the presence of queuosine in RNA. Application of polyacrylamide containing a covalently bound boronic acid (original) (raw)
Journal Article
Institut für Biologie III der Universität Freiburg
Schänzlestr. 1, D-7800 Freiburg, FRG
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Institut für Biologie III der Universität Freiburg
Schänzlestr. 1, D-7800 Freiburg, FRG
Search for other works by this author on:
Revision received:
09 September 1985
Accepted:
09 September 1985
Published:
11 October 1985
PDF Cite
Gabar L. Igloi, Hans Kössel, Affinity electrophoresis for monitoring terminal phosphorylation and the presence of queuosine in RNA. Application of polyacrylamide containing a covalently bound boronic acid, Nucleic Acids Research, Volume 13, Issue 19, 11 October 1985, Pages 6881–6898, https://doi.org/10.1093/nar/13.19.6881
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Abstract
An affinity electrophoretic method has been developed to study the state of terminal phosphorylation of RNAs and the presence of the hypermodified base Q in tRNA. It is based on the copolymerization of acryloylaminophenylboronic acid into standard polyacrylamide gels and the interaction of this derivative with free cis-diol groups present in the RNA. In the case of terminal phosphorylation, free ribose groups are present either as such, or may be introduced by enzymatic reactions specific for a particular phosphorylation pattern (e.g. using T4 RNA ligase or guanylyltransferase). Additionally, tRNA species containing the Q base may be resolved from Q-lacking tRNAs by boronate affinity electrophoresis. The introduction of a non-destructive, one-step electrophoretic procedure not only offers an alternative to classical analytical methods, but also provides a means of isolating such populations of RNAs for which other methods are unavailable or are less convenient.
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