BMB Reports
- Volume 31 Issue 2
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- Pages.177-182
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- 1998
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- 1976-670X(eISSN)
In Vitro Selection of Hammerhead Ribozymes with Optimized Stems I and III
- Sim, So-Yeong (Department of Chemistry, Korea Advanced Institute of Science and Technology) ;
- Kim, Se-Mi (Department of Chemistry, Korea Advanced Institute of Science and Technology) ;
- Kim, Ha-Dong (Department of Chemistry, Seoul National University) ;
- Ahn, Jeong-Keun (Department of Microbiology, Chungnam National University) ;
-
Lee, Young-Hoon
(Department of Chemistry, Korea Advanced Institute of Science and Technology)
;
- Cho, Bong-Rae (Department of Chemistry, Chongju University) ;
- Park, In-Won (Department of Chemistry, Seoul National University)
- Received : 1997.11.22
- Published : 1998.03.31
Abstract
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.