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6 results on '"Masayoshi Hayami"'

1. A Concept for Selection of Codon-Suppressor tRNAs Based on Read-Through Ribosome Display in an In Vitro Compartmentalized Cell-Free Translation System

Author

Atsushi Ogawa, Masayoshi Hayami, Shinsuke Sando, and Yasuhiro Aoyama

Subjects
Genetics, QH426-470, Biochemistry, QD415-436
Abstract

Here is presented a concept for in vitro selection of suppressor tRNAs. It uses a pool of dsDNA templates in compartmentalized water-in-oil micelles. The template contains a transcription/translation trigger, an amber stop codon, and another transcription trigger for the anticodon- or anticodon loop-randomized gene for tRNASer. Upon transcription are generated two types of RNAs, a tRNA and a translatable mRNA (mRNA-tRNA). When the tRNA suppresses the stop codon (UAG) of the mRNA, the full-length protein obtained upon translation remains attached to the mRNA (read-through ribosome display) that contains the sequence of the tRNA. In this way, the active suppressor tRNAs can be selected (amplified) and their sequences read out. The enriched anticodon (CUA) was complementary to the UAG stop codon and the enriched anticodon-loop was the same as that in the natural tRNASer.

Published
2012
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2. In vitro selection of RNA aptamer against Escherichia coli release factor 1

Author

Atsushi Ogawa, Shinsuke Sando, Teruyuki Nishi, Masayoshi Hayami, and Yasuhiro Aoyama

Subjects
Aptamer, Clinical Biochemistry, Drug Evaluation, Preclinical, Pharmaceutical Science, Biochemistry, Substrate Specificity, Structure-Activity Relationship, Drug Discovery, Genes, Suppressor, Molecular Biology, Binding Sites, Chemistry, Escherichia coli Proteins, Organic Chemistry, Peptide Termination Factors, RNA, Aptamers, Nucleotide, Molecular biology, Genetic translation, Stop codon, Terminator (genetics), Transfer RNA, Codon, Terminator, Molecular Medicine, Release factor
Abstract

A pool of 84-nt RNAs containing a randomized sequence of 50 nt was selected against gel-immobilized Escherichia coli release factor 1 (RF-1) responsible for translation termination at amber (UAG) stop codon. The strongest aptamer (class II-1) obtained from 43 clones bound to RF-1, but not to UAA/UGA-targeting RF-2, with Kd = 30+/-6 nM (SPR). A couple of unpaired hairpin domains in the aptamer were suggested as the sites of attachment of RF-1. By binding to and hence inhibiting the action of RF-1 specifically or bio-orthogonally, aptamer class II-1 enhanced the amber suppression efficiency in the presence of an anticodon-adjusted (CUA) suppressor tRNA without practically damaging the protein translation machinery of the cell-free extract of E. coli, as confirmed by the translation of amber-mutated (gfp(amber141) or gfp(amber178)) and wild-type (gfp(wild)) genes of GFP.

Published
2007
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3. Transcription monitoring using fused RNA with a dye-binding light-up aptamer as a tag: a blue fluorescent RNA

Author

Shinsuke Sando, Yasuhiro Aoyama, Atsushi Narita, and Masayoshi Hayami

Subjects
Transcription, Genetic, Dye binding, Aptamer, Catalysis, Fluorescence, Transcription (biology), Materials Chemistry, heterocyclic compounds, neoplasms, Chemistry, fungi, Metals and Alloys, food and beverages, RNA, General Chemistry, Aptamers, Nucleotide, Molecular biology, Surfaces, Coatings and Films, Electronic, Optical and Magnetic Materials, Biochemistry, Ceramics and Composites, Nucleic Acid Conformation, biological phenomena, cell phenomena, and immunity, Light Up, Fluorescent tag
Abstract

The "light-up" RNA aptamer-Hoechst pair can be used as a fluorescent tag to monitor transcription processes.

Published
2008

4. A Concept for Selection of Codon-Suppressor tRNAs Based on Read-Through Ribosome Display in an In Vitro Compartmentalized Cell-Free Translation System

Author

Yasuhiro Aoyama, Shinsuke Sando, Masayoshi Hayami, and Atsushi Ogawa

Subjects
Genetics, Messenger RNA, Article Subject, lcsh:QH426-470, Biology, Biochemistry, Stop codon, law.invention, lcsh:Biochemistry, Open reading frame, lcsh:Genetics, law, Transcription (biology), Transfer RNA, Ribosome display, Suppressor, lcsh:QD415-436, Molecular Biology, Gene, Research Article
Abstract

Here is presented a concept forin vitroselection of suppressor tRNAs. It uses a pool of dsDNA templates in compartmentalized water-in-oil micelles. The template contains a transcription/translation trigger, an amber stop codon, and another transcription trigger for the anticodon- or anticodon loop-randomized gene for tRNASer. Upon transcription are generated two types of RNAs, a tRNA and a translatable mRNA (mRNA-tRNA). When the tRNA suppresses the stop codon (UAG) of the mRNA, the full-length protein obtained upon translation remains attached to the mRNA (read-through ribosome display) that contains the sequence of the tRNA. In this way, the active suppressor tRNAs can be selected (amplified) and their sequences read out. The enriched anticodon (CUA) was complementary to the UAG stop codon and the enriched anticodon-loop was the same as that in the natural tRNASer.

Published
2012

5. Tuning the Hoechst Dye into Color-changing Fluorescent pH Indicator in an Acidic Range

Author

Shinsuke Sando, Yasuhiro Aoyama, Masayoshi Hayami, and Atsushi Narita

Subjects
endocrine system, Range (particle radiation), chemistry.chemical_compound, animal structures, chemistry, pH indicator, heterocyclic compounds, General Chemistry, biological phenomena, cell phenomena, and immunity, Photochemistry, neoplasms, Fluorescence
Abstract

We developed a new fluorescent pH indicator from a common Hoechst 33258 fluorescent dye. Unlike the parent compound, the improved Hoechst probe, wherein two methyl substituents were introduced into...

Published
2008
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