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Guide DNA technique in bacterial ribonuclease P reaction for effective processing of tRNA precursor
Author(s) -
Tanaka Terumichi,
Hori Yoshiaki,
Kikuchi Yo
Publication year - 2002
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1042/ba20020047
Subject(s) - cleavage (geology) , ribonuclease , dna , transfer rna , biology , biochemistry , microbiology and biotechnology , rna , gene , paleontology , fracture (geology)
Previously, we found that a small (approx. 20‐mer) DNA hybridizing to the 5′‐leader region of a tRNA precursor enhances the cleavage efficiency in bacterial ribonuclease P reaction. We named this technique the ‘guide DNA technique’. Detailed analyses showed that the length of the guide DNA, concentration of the guide DNA and the hybridizing position affected the cleavage efficiency: for an effective cleavage reaction, guide DNA should be designed to hybridize to the region on the cleavage site, should be 20 bases or more in length and should be of high concentration. The presence of a 5′‐flanking region in the DNA did not affect the cleavage reaction. The guide DNA technique is a useful tool for effective preparation of mature tRNA molecules in vitro .