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

Identification of oligomerizing peptides.

A Dhiman1, M E Rodgers, R Schleif

  • 1Biology Department, Johns Hopkins University, Baltimore, Maryland 21218, USA.

The Journal of Biological Chemistry
|March 29, 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 conundrum of olecranon aperture and its relation to the distal end of the humerus in a modern Indian population: An anatomical and surgical perspective.

Morphologie : bulletin de l'Association des anatomistes·2022
Same author

Synthesis and characterization of arabinoxylan-bis[2-(methacryloyloxy)ethyl] phosphate crosslinked copolymer network by high energy gamma radiation for use in controlled drug delivery applications.

International journal of biological macromolecules·2022
Same author

Synchronous carcinoma breast with chronic myelogenous leukemia: a rare presentation.

Indian journal of cancer·2010
Same author

Data-driven computer simulation of human cancer cell.

Annals of the New York Academy of Sciences·2004
Same author

Gonorrhoea in a south London genitourinary medicine department.

International journal of STD & AIDS·2003
Same author

The child patient and consent to treatment: legal overview.

British journal of community nursing·2002

Researchers identified functional chimeric proteins by linking random peptides to the AraC DNA binding domain. This method enabled the dimerization of the AraC domain, activating transcription from arabinose operon promoters.

Area of Science:

  • Molecular Biology
  • Genetics
  • Protein Engineering

Background:

  • The AraC DNA binding domain requires dimerization to activate transcription of the arabinose operon.
  • Monomeric AraC DNA binding domain is transcriptionally inactive.

Purpose of the Study:

  • To identify novel peptide sequences that can induce dimerization of the AraC DNA binding domain.
  • To develop a method for discovering chimeric proteins with engineered dimerization properties.

Main Methods:

  • Construction of a high-diversity plasmid library by inserting random 90-base oligonucleotides upstream of the AraC DNA binding domain gene.
  • Transformation of expression vectors and selection of functional chimeric proteins based on growth in minimal arabinose medium.
  • Purification and characterization of candidate proteins, including equilibrium analytical ultracentrifugation.

Related Experiment Videos

Main Results:

  • A library of chimeric proteins was generated, with several candidates showing arabinose operon activation (Ara+).
  • One purified chimeric protein demonstrated dimerization with a dissociation constant (Kd) of approximately 2 micromolar.
  • The study successfully identified peptide additions that promote AraC DNA binding domain dimerization.

Conclusions:

  • Engineered peptide sequences can effectively induce dimerization of the AraC DNA binding domain.
  • This approach provides a powerful tool for discovering and engineering protein-protein interactions and functional domains.
  • The identified chimeric proteins have potential applications in synthetic biology and gene regulation systems.