Sequence‐specific cleavage of RNA by a hybrid ribonuclease H

FEBS Letters

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