In vitro selection of hammerhead ribozymes with optimized stems I and III

A pool of cis-acting hammerhead ribozymes randomized in their substrate recognition sequences was constructed. A variety of active cis-acting ribozymes which had various structures of stems I and III was selected from the pool by in vitro selection. The selected ribozymes were cloned and sequenced, The relationship between the cleavage efficiency and base-pairing in stems I and III of the selected ribozymes was investigated. The ribozymes with the smaller difference in folding energies between the active conformation and the stable but inactive conformation showed a tendency to have the better cleavage efficiency. The optimum length of stem I was 5 or 6 bases while the longer stem III, in general, appeared to be required for efficient cleavage. The specificity of the ribozyme reaction is discussed in terms of the length of stems I and III.
Publisher
SPRINGER-VERLAG SINGAPORE PTE LTD
Issue Date
1998-03
Language
ENG
Keywords

HEPATITIS-B VIRUS; CLEAVAGE RATES; RNA; SITE

Citation

JOURNAL OF BIOCHEMISTRY AND MOLECULAR BIOLOGY, v.31, no.2, pp.177 - 182

ISSN
1225-8687
URI
http://hdl.handle.net/10203/73610
Appears in Collection
CH-Journal Papers(저널논문)
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