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Sequence‐specific cleavage of RNA by a hybrid ribonuclease H
Author(s) -
Nakai Chieko,
Konishi Aritomo,
Komatsu Yasuo,
Inoue Hideo,
Ohtsuka Eiko,
Kanaya Shigenori
Publication year - 1994
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(94)80386-2
Subject(s) - phosphodiester bond , rna , rnase p , rnase h , ribonuclease , cleavage (geology) , enzyme , ribonuclease t1 , biochemistry , dna , base pair , biology , microbiology and biotechnology , chemistry , gene , paleontology , fracture (geology)
Site‐specific cleavage of the 22‐, 132‐ and 534‐base RNAs by the DNA/protein hybrid R Nase H were examined. The 22‐base RNA was chemically synthesized, and 132‐ and 534‐base RNAs were prepared by run‐off transcription. The hybrid enzyme cleaves these RNAs, which contain a single target sequence, primarily at the unique phosphodiester bond within the target sequence. The hybrid enzyme performs multiple turnovers, and at a substrate/enzyme ratio of 10:1 the RNAs are almost completely cleaved by the hybrid enzyme at 37°C within 1 h. We propose that hybrid RNase H molecules with various oligodeoxyribonucleotides function as RNA restriction enzymes and are useful for structural and functional studies of RNA.