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

The bacteriophage 434 operator/repressor system in yeast

C I Webster1, W J Brammar

  • 1ICI/University Joint Laboratory, Department of Biochemistry, University of Leicester, UK.

Microbiology (Reading, England)
|September 1, 1995
PubMed
Summary

The bacteriophage 434 operator/repressor system can regulate gene expression in yeast. However, a modified repressor variant showed toxicity, hindering its application in eukaryotic gene regulation.

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 activity of a single-stranded promoter of plasmid ColIb-P9 depends on its secondary structure.

Molecular microbiology·2004
Same author

HMG-1 enhances HMG-I/Y binding to an A/T-rich enhancer element from the pea plastocyanin gene.

European journal of biochemistry·2001
Same author

Kinetic analysis of high-mobility-group proteins HMG-1 and HMG-I/Y binding to cholesterol-tagged DNA on a supported lipid monolayer.

Nucleic acids research·2000
Same author

Co-localization of the inwardly rectifying potassium ion channel, Kir2.2, and the substance P receptor in single locus coeruleus neurons.

Annals of the New York Academy of Sciences·2000
Same author

Characterisation of Kir2.0 proteins in the rat cerebellum and hippocampus by polyclonal antibodies.

Histochemistry and cell biology·2000
Same author

Expression of leading region genes on IncI1 plasmid ColIb-P9: genetic evidence for single-stranded DNA transcription.

Microbiology (Reading, England)·1999

Area of Science:

  • Molecular Biology
  • Microbiology
  • Genetics

Background:

  • Bacteriophage operator/repressor systems are crucial for controlling gene expression in prokaryotes.
  • Investigating the functionality of these systems in eukaryotic hosts can reveal conserved regulatory mechanisms and potential biotechnological applications.

Purpose of the Study:

  • To determine if the bacteriophage 434 operator/repressor system can function within a eukaryotic cell, specifically Saccharomyces cerevisiae.
  • To assess the impact of operator placement on gene repression efficiency.
  • To evaluate the utility of a modified repressor with altered binding specificity.

Main Methods:

  • Insertion of an idealized 434 operator sequence into various locations of the Saccharomyces cerevisiae PGK promoter.
  • Linking modified promoters to a beta-galactosidase reporter gene.

Related Experiment Videos

  • Expression of the 434 cI repressor gene from a 2-micron plasmid.
  • Assessing gene expression levels and cell growth.

    • Main Results:

    • Significant repression of beta-galactosidase expression was observed when the 434 operator was integrated into the PGK promoter at different positions.
    • The bacteriophage 434 repressor effectively controlled gene expression in yeast.
    • A variant repressor (434P22) with P22 repressor binding specificity exhibited severe toxicity, inhibiting yeast cell growth.

    Conclusions:

    • The bacteriophage 434 operator/repressor system is functional in Saccharomyces cerevisiae, demonstrating cross-kingdom regulatory potential.
    • The placement of the operator sequence influences repression efficiency.
    • The tested 434P22 repressor variant is unsuitable for eukaryotic applications due to growth inhibition.