Effective structure of a leader open reading frame for enhancing the expression of GC-rich genes.

Effective structure of a leader open reading frame for enhancing the expression of GC-rich genes. Research Abstract Details 

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Effective structure of a leader open reading frame for enhancing the expression of GC-rich genes. Abstract Text:

Masami Ishida,Tairo Oshima,

To overexpress broad kinds of GC-rich genes in Escherichia coli, we examined how the structures of leader open reading frames (leader ORFs) affect the expression of GC-rich genes, such as polA, trpA, and trpB, from Thermus thermophilus. When a leader ORF overlapped with the polA-initiation codon by 1 bp in the TGATG motif, gene expression increased by more than 3-fold compared to when a leader ORF was several-bp distant from the initiation codon. A 4-bp overlap with the ATGA motif was more effective than a 1-bp overlap with the TGATG motif. When a 4-bp overlapping leader ORF was placed in front of the successive trpB and trpA genes, the trpA gene was poorly expressed whereas the trpB gene was overexpressed. Mutation analysis revealed that the expression of the trpA gene was strongly enhanced by replacing G and C in the translation termination region of the leader ORF with A and T. In contrast, other mutations, such as alterations between synonymous codons in the trpA-coding region, produced diminished gene expression. Using the most effective leader ORF obtained from these results, new expression vectors were constructed.

Effective structure of a leader open reading frame for enhancing the expression of GC-rich genes. Publishing Authors By Initials

M Ishida,T Oshima,

For similar enzymes and coenzymes: enzymes: lyases: carbon-oxygen lyases: hydro-lyases: tryptophan synthase research abstracts see: enzymes and coenzymes: enzymes: lyases: carbon-oxygen lyases: hydro-lyases: tryptophan synthase research

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Effective structure of a leader open reading frame for enhancing the expression of GC-rich genes. Journal Published:

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

Journal: Journal of biochemistry

VOLUME: 132

Page Numbers: 63-70

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: Jul

YEAR: 2002

Effective structure of a leader open reading frame for enhancing the expression of GC-rich genes. Information

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LANGUAGE: eng

NlmUniqueID: 376600

Effective structure of a leader open reading frame for enhancing the expression of GC-rich genes. Keywords Mesh Terms:

KEYWORDS: Tryptophan Synthase

MESH TERMS: biosynthesis

Chemical & Substance for Abstract: Effective structure of a leader open reading frame for enhancing the expression of GC-rich genes. Information

Substance Name: Tryptophan Synthase

Registry Number: EC 4.2.1.20

Grant and Affiliation Information for Effective structure of a leader open reading frame for enhancing the expression of GC-rich genes.

AFFILIATION: Laboratory of Marine Biochemistry, Tokyo University of Fisheries, 4-5-7 Konan, Minato-ku, Tokyo 108-8477, Japan. ishida@tokyo-u-fish.ac.jp

Country: Japan

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MEDLINETA: J Biochem

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ACCESSION NUMBER: AB049969

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