Guide DNA technique in bacterial ribonuclease P reaction for effective processing of tRNA precursor

Terumichi Tanaka; Yoshiaki Hori; Yo Kikuchi

doi:10.1042/BA20020047

Guide DNA technique in bacterial ribonuclease P reaction for effective processing of tRNA precursor

Terumichi Tanaka, Yoshiaki Hori, Yo Kikuchi

Research output: Contribution to journal › Article › peer-review

4 Citations (Scopus)

Abstract

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.

Original language	English
Pages (from-to)	85-88
Number of pages	4
Journal	Biotechnology and Applied Biochemistry
Volume	36
Issue number	2
DOIs	https://doi.org/10.1042/BA20020047
Publication status	Published - 2002 Oct
Externally published	Yes

Keywords

Escherichia coli
Leader sequence
Maturation
Ribozyme
RNase P

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)
Biochemistry
Biotechnology

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10.1042/BA20020047

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title = "Guide DNA technique in bacterial ribonuclease P reaction for effective processing of tRNA precursor",

abstract = "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.",

keywords = "Escherichia coli, Leader sequence, Maturation, Ribozyme, RNase P",

author = "Terumichi Tanaka and Yoshiaki Hori and Yo Kikuchi",

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T1 - Guide DNA technique in bacterial ribonuclease P reaction for effective processing of tRNA precursor

AU - Tanaka, Terumichi

AU - Hori, Yoshiaki

AU - Kikuchi, Yo

PY - 2002/10

Y1 - 2002/10

N2 - 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.

AB - 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.

KW - Escherichia coli

KW - Leader sequence

KW - Maturation

KW - Ribozyme

KW - RNase P

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