Use of RNase P for efficient preparation of yeast tRNATyr transcript and its mutants.

Use of RNase P for efficient preparation of yeast tRNATyr transcript and its mutants. Research Abstract Details 

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Use of RNase P for efficient preparation of yeast tRNATyr transcript and its mutants. Abstract Text:

Jun-Ichi Fukunaga,Masaki Gouda,Koji Umeda,Satoshi Ohno,Takashi Yokogawa,Kazuya Nishikawa,

Because T7 RNA polymerase has a strong preference for particular sequences to initiate transcription, some RNAs having pyrimidine-rich sequences at their 5'-end (yeast tRNA(Tyr), for example) are hardly transcribed by this enzyme. To circumvent this inconvenience, we have developed an efficient method for in vitro preparation of such tRNAs. The RNA of interest is first transcribed as a precursor form that has purine-rich extra sequences at its 5'-end, then processed with RNase P to generate the objective tRNAs. By using this protocol, we were able to prepare easily and efficiently yeast tRNA(Tyr) transcript and its mutants harboring base substitutions within the anticodon loop and/or acceptor stem regions. Aminoacylation analyses of these tRNA transcripts with yeast tyrosyl-tRNA synthetase revealed that the replacement of G34 by C34 (mutation to amber suppressor) severely impaired the aminoacylation, whereas the replacement of the U4:G69 wobble base-pair in the acceptor stem region by C4:G69 normal Watson-Crick type base-pair improved it.

Use of RNase P for efficient preparation of yeast tRNATyr transcript and its mutants. Publishing Authors By Initials

J Fukunaga,M Gouda,K Umeda,S Ohno,T Yokogawa,K Nishikawa,

For similar genetic processes: gene expression: transcription, genetic research abstracts see: genetic processes: gene expression: transcription, genetic research

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Use of RNase P for efficient preparation of yeast tRNATyr transcript and its mutants. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of biochemistry

VOLUME: 139

Page Numbers: 123-7

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: Jan

YEAR: 2006

Use of RNase P for efficient preparation of yeast tRNATyr transcript and its mutants. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 376600

Use of RNase P for efficient preparation of yeast tRNATyr transcript and its mutants. Keywords Mesh Terms:

KEYWORDS: Transcription, Genetic

MESH TERMS: metabolism

Chemical & Substance for Abstract: Use of RNase P for efficient preparation of yeast tRNATyr transcript and its mutants. Information

Substance Name: Ribonuclease P

Registry Number: EC 3.1.26.5

Grant and Affiliation Information for Use of RNase P for efficient preparation of yeast tRNATyr transcript and its mutants.

AFFILIATION: Department of Biomolecular Science, Faculty of Engineering, Gifu University, Yanagido 1-1, Gifu 501-1193.

Country: Japan

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MEDLINETA: J Biochem

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