Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Protein-protein interaction panel using mouse full-length cDNAs.

H Suzuki1, Y Fukunishi, I Kagawa

  • 1Laboratory for Genome Exploration Research Group, RIKEN Genomic Sciences Center, Yokohama 230-0045, Japan.

Genome Research
|October 10, 2001
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

The transcriptional landscape of the mammalian genome.

Science (New York, N.Y.)·2005
Same author

Analysis of the mouse transcriptome based on functional annotation of 60,770 full-length cDNAs.

Nature·2002
Same author

Anti-alpha-fodrin antibodies in Sjögren's syndrome in children.

The Journal of rheumatology·2001
Same author

Production of neuropeptide substance P by synovial fibroblasts from patients with rheumatoid arthritis and osteoarthritis.

Neuroscience letters·2001
Same author

An investigation of cell proliferation and soluble mediators induced by interleukin 1beta in human synovial fibroblasts: comparative response in osteoarthritis and rheumatoid arthritis.

Inflammation research : official journal of the European Histamine Research Society ... [et al.]·2001
Same author

Urinary prostaglandin D synthase (beta-trace) excretion increases in the early stage of diabetes mellitus.

Nephron·2001

This study introduces a novel, high-throughput assay for analyzing protein-protein interactions (PPIs) using PCR and a mammalian two-hybrid system. The method efficiently identifies numerous PPIs, aiding in understanding protein functions and biological processes.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genomics

Background:

  • Protein-protein interactions (PPIs) are crucial for cellular functions.
  • Systematic analysis of PPIs is essential for understanding complex biological pathways.
  • Existing methods for PPI analysis can be time-consuming and low-throughput.

Purpose of the Study:

  • To develop a novel, high-throughput assay system for systematic analysis of protein-protein interactions.
  • To enable rapid sample preparation using PCR-mediated methods.
  • To facilitate the discovery of novel PPIs and functional insights into uncharacterized proteins.

Main Methods:

  • Developed a high-throughput assay system based on the mammalian two-hybrid method.
  • Utilized PCR-mediated rapid sample preparation with gene-specific primers.

Related Experiment Videos

  • Implemented a semiautomatic multiple dispenser system for 384-well plates, enabling over 20,000 assays daily.

    • Main Results:

    • Successfully constructed assay samples up to 3.6 kb using two rounds of PCR.
    • Detected 145 protein-protein interactions in a pilot study of 3500 samples from mouse cDNAs.
    • Identified significant interaction clusters and predicted functions for uncharacterized proteins.

    Conclusions:

    • The developed high-throughput assay system enables efficient and systematic analysis of PPIs.
    • This system, combined with a comprehensive cDNA clone bank, can accelerate the discovery of PPIs in mice.
    • Facilitates understanding of uncharacterized protein functions and molecular mechanisms of biological processes.