FEBS Letters | |
Sequence‐specific cleavage of RNA by a hybrid ribonuclease H | |
Kanaya, Shigenori1  Komatsu, Yasuo2  Inoue, Hideo2  Nakai, Chieko1  Ohtsuka, Eiko2  Konishi, Aritomo2  | |
[1] Protein Engineering Research Institute, 6-2-3, Furuedai, Suita, Osaka 565, Japan;Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo 060, Japan | |
关键词: RNase H; Hybrid enzyme; Sequence specificity; Protein engineering; RNase H; ribonuclease H; | |
DOI : 10.1016/0014-5793(94)80386-2 | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
【 摘 要 】
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.
【 授权许可】
Unknown
【 预 览 】
Files | Size | Format | View |
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RO201912020299152ZK.pdf | 639KB | download |