NDLI: Kinetic characterization of intramolecular and intermolecular hammerhead RNAs with stem II deletions.

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Kinetic characterization of intramolecular and intermolecular hammerhead RNAs with stem II deletions.

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Kinetic characterization of intramolecular and intermolecular hammerhead RNAs with stem II deletions.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Long, D. M. Uhlenbeck, O. C.
Description	Author Affiliation: Long DM ( Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309-0215.);
Abstract	A method is described to obtain intramolecular cleavage rates for the hammerhead ribozyme during in vitro transcription. By avoiding RNA purification and renaturation, the potential for formation of inactive conformations of the RNA is minimized. By showing that an intramolecular hammerhead and a closely related intermolecular hammerhead cleave at the same rate under a given set of conditions, we confirm that both reactions probably have the same rate-limiting step. An in vitro selection strategy was used to isolate active hammerheads from a library of molecules where six randomized nucleotides replaced stem-loop II. The sequence and number of nucleotides which replace stem-loop II have large effects on hammerhead cleavage activity. The relative activities of three sequences selected from the intramolecular library are the same when the sequences are transferred into an intermolecular hammerhead background.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	15
Volume Number	91
Language	English
Publisher	National Academy of Sciences
Publisher Date	1994-08-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	RNA, Catalytic Metabolism Genetic Techniques Kinetics Molecular Sequence Data Nucleic Acid Conformation Polymerase Chain Reaction Chemistry Sequence Deletion Transcription, Genetic Research Support, U.S. Gov't, P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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