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 Video

Updated: Jun 17, 2026

Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens
08:49

Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens

Published on: June 6, 2020

Screening libraries to identify proteins with desired binding activities using a split-GFP reassembly assay.

Meredith E Jackrel1, Aitziber L Cortajarena, Tina Y Liu

  • 1Departments of Chemistry, Yale University, New Haven, Connecticut 06520, USA.

ACS Chemical Biology
|December 30, 2009
PubMed
Summary

Researchers developed a novel split-green fluorescent protein (GFP) assay to efficiently screen protein libraries for specific binding properties. This method identifies designer proteins with high accuracy and ease of automation.

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

Engineered protein nanoclusters reduce liver fibrosis and hepatocellular carcinoma in mice models.

Bioactive materials·2026
Same author

High-throughput screening approach identifies substrate-selective Hsp104 variants that counter amyloid seeding with diminished off-target effects.

Molecular cell·2026
Same author

Programmable kinetic barcoding for multiplexed RNA detection with Cas13a.

Nature biomedical engineering·2026
Same author

epiGPTope: A Machine Learning-Based Epitope Generator and Classifier.

ACS synthetic biology·2026
Same author

Substituent-Based Modulation of Self-Assembly and Immunogenicity of Amphipathic Peptides.

Advanced science (Weinheim, Baden-Wurttemberg, Germany)·2026
Same author

Magnetic nanoparticles as transducers for quick and direct virus detection in clinical samples.

Nanoscale·2025

Area of Science:

  • Protein engineering
  • Biotechnology
  • Molecular biology

Background:

  • Designer proteins with specific target-binding capabilities are crucial for various applications.
  • Protein libraries are commonly screened to discover novel binding proteins.
  • Existing screening methods can be complex or prone to false positives.

Purpose of the Study:

  • To describe the construction of a naive protein library.
  • To present a split-green fluorescent protein (GFP) reassembly assay for screening protein libraries.
  • To identify proteins with novel binding properties using this assay.

Main Methods:

  • Construction of a naive protein library.
  • Utilizing a split-GFP reassembly assay for high-throughput screening.

More Related Videos

Semi-automated Biopanning of Bacterial Display Libraries for Peptide Affinity Reagent Discovery and Analysis of Resulting Isolates
13:49

Semi-automated Biopanning of Bacterial Display Libraries for Peptide Affinity Reagent Discovery and Analysis of Resulting Isolates

Published on: December 6, 2017

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli
08:46

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

Published on: January 6, 2015

Related Experiment Videos

Last Updated: Jun 17, 2026

Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens
08:49

Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens

Published on: June 6, 2020

Semi-automated Biopanning of Bacterial Display Libraries for Peptide Affinity Reagent Discovery and Analysis of Resulting Isolates
13:49

Semi-automated Biopanning of Bacterial Display Libraries for Peptide Affinity Reagent Discovery and Analysis of Resulting Isolates

Published on: December 6, 2017

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli
08:46

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

Published on: January 6, 2015

  • Screening the library against specific peptide sequences.

    • Main Results:

    • The split-GFP assay demonstrated simplicity, robustness, and ease of automation.
    • The assay effectively identified proteins with specific binding properties.
    • Absence of false positives was a key advantage of the split-GFP method.

    Conclusions:

    • The split-GFP reassembly assay is a powerful and reliable tool for discovering designer proteins with specific binding functions.
    • This method facilitates the identification of novel protein binders for diverse applications.
    • The described approach offers an efficient and automatable solution for protein library screening.