A new approach to gene mutation analysis using "GFP-Display".

A new approach to gene mutation analysis using "GFP-Display". Research Abstract Details 

Research Abstract Table of Contents

Jump to the:

A new approach to gene mutation analysis using "GFP-Display". Abstract Text:

T Aoki,R Ami,H Onagi,H Fujino,H Watabe,

The unique behavior of green fluorescent protein (GFP) on SDS-PAGE was applied to the detection of a single amino acid substitution in GFP-tagged polypeptides. This simple detection method using SDS/urea gels was designated GFP-display. The N-terminal 18 or 37 amino acids of K-Ras was used as a model GFP-tagged polypeptide. K-ras exon 1 was fused to a gfp cDNA at each end and expressed in Escherichia coli. Amino acid number 12 of K-Ras (wild type; Gly) was changed to Ser, Arg, Cys, Asp, Ala, or Val, and the mobility shift of the greenish fluorescent bands in the SDS/urea gel was analyzed. These mutants were easily detected by GFP-display; however, detection depended strongly on the urea concentration and electrophoresis temperature. Subsequently, GFP-display was applied to the 36 amino acids encoding human p53 exon 7. Amino acid number 248 (wild type; Arg) was changed to Gly, Trp, Gln, Pro, or Leu, and similar mobility shifts were observed. GFP-display could be coupled with an in vitro translation system. Fluorescent active GFP and GFP-Ras fusion proteins were synthesized within a few hours. GFP-display shows potential as a modern approach to gene mutation analysis at the protein level, and is a useful method for protein engineering studies.

A new approach to gene mutation analysis using "GFP-Display". Publishing Authors By Initials

T Aoki,R Ami,H Onagi,H Fujino,H Watabe,

For similar organic chemicals: urea research abstracts see: organic chemicals: urea research

PUBMED ID PMID:

MEDLINE DATE:

A new approach to gene mutation analysis using "GFP-Display". Journal Published:

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

Journal: Journal of biochemistry

VOLUME: 127

Page Numbers: 627-33

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: Apr

YEAR: 2000

A new approach to gene mutation analysis using "GFP-Display". Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 376600

A new approach to gene mutation analysis using "GFP-Display". Keywords Mesh Terms:

KEYWORDS: Urea

MESH TERMS: genetics

Chemical & Substance for Abstract: A new approach to gene mutation analysis using "GFP-Display". Information

Substance Name: Urea

Registry Number: 57-13-6

Grant and Affiliation Information for A new approach to gene mutation analysis using "GFP-Display".

AFFILIATION: Department of Biochemistry, Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido, Kanazawa, Ishikari-Tobetsu, Hokkaido 061-0293, Japan. aokit@hoku-iryo-u.ac.jp

Country: JAPAN

AGENCY:

GRANT:

ACRONYM:

MEDLINETA: J Biochem

REFSOURCE:

DATABASENAME:

ACCESSION NUMBER:

Number Hits: 0

A new approach to gene mutation analysis using "GFP-Display" Related Publications

• A new approach to gene mutation analysis using "GFP-Display".
• A new enzyme, NADPH-dihydropteridine reductase in bovine liver.
• Absolute quantitation of myocardial blood flow with (201)Tl and dynamic SPECT in canine: optimisation and validation of kinetic modelling.
• Actin-binding specificity of marine macrolide toxins, mycalolide B and kabiramide D.
• Antibody against a novel, myriocin (ISP-I)-based immunosuppressant, FTY720.
• CYR61 is a novel gene associated with temperature-dependent changes in fish metabolism as revealed by cDNA microarray analysis on a medaka Oryzias latipes cell line.
• Detection of blotted antigen using a fusion protein between protein A and pepsinogen C.
• Differential scanning calorimetry of light meromyosin fragments having various lengths of carp fast skeletal muscle isoforms.
• Direct evidence for redundant segmental replacement between multiple 18S rRNA genes in a single Prototheca strain.
• Divergent evolution of the myosin heavy chain gene family in fish and tetrapods: evidence from comparative genomic analysis.
• Expression, Localization and Modification of YxeE Spore Coat Protein in Bacillus subtilis.
• Fatty acid synthase gene is up-regulated by hypoxia via activation of Akt and sterol regulatory element binding protein-1.
• Ligand-dependent transcriptional activities of four torafugu pufferfish Takifugu rubripes peroxisome proliferator-activated receptors.
• Lower activation energy for sliding of F-actin on a less thermostable isoform of carp myosin.
• Modification of GerQ reveals a functional relationship between Tgl and YabG in the coat of Bacillus subtilis spores.
• Molecular Characterization of Japanese Sillago Vitellogenin and Changes in Its Expression Levels on Exposure to 17beta-Estradiol and 4-tert-Octylphenol.
• Molecular cloning and functional characterization of crustapain: a distinct cysteine proteinase with unique substrate specificity from northern shrimp Pandalus borealis.
• Myocyte enhancer factor 2 regulates expression of medaka Oryzias latipes fast skeletal myosin heavy chain genes in a temperature-dependent manner.
• Myosin subfragment-1 isoforms having different heavy chain structures from fast skeletal muscle of thermally acclimated carp.
• Relationship between postmortem changes and browning of boiled, dried, and seasoned product made from Japanese common squid (Tedarodes pacificus) mantle muscle.
• RhoC Promotes Metastasis via Activation of the Pyk2 Pathway in Prostate Cancer.
• Role of the interventional radiologist in treating obstetric-gynecologic pathology.
• Synthesis and characterization of the spore proteins of Bacillus subtilis YdhD, YkuD, and YkvP, which carry a motif conserved among cell wall binding proteins.
• Tumor-endothelial cell interactions: Therapeutic potential.
• Ultrastructural and histochemical aspects of zinc accumulation by fish scales.
• Unphosphorylated twitchin forms a complex with actin and myosin that may contribute to tension maintenance in catch.
• [Illness, hospitalization and children]
• [Proximal axonal injuries as experimental models for adult motoneuron degeneration]
• cDNA cloning and characterization of temperature-acclimation-associated light meromyosins from grass carp fast skeletal muscle.