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

Regulated expression by readthrough translation from a plasmid-encoded beta-galactosidase.

P O'Farrell, B Polisky, D H Gelfand

    Journal of Bacteriology
    |May 1, 1978
    PubMed
    Summary

    This study characterizes beta-galactosidase expression from plasmid pBGP120, finding that inserting DNA fragments at the EcoRI site creates stable, fused proteins. Expression levels are high and regulated by cyclic AMP and lac repressor.

    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

    Adjustment to Heart Disease: Helping families cope.

    Canadian family physician Medecin de famille canadien·2011
    Same author

    DNA-dependent DNA polymerases.

    Current protocols in molecular biology·2008
    Same author

    Use of a thin film biosensor for rapid visual detection of PCR products in a multiplex format.

    Biosensors & bioelectronics·2001
    Same author

    Silicon-based biosensors for rapid detection of protein or nucleic acid targets.

    Clinical chemistry·2001
    Same author

    Rapid multiserotype detection of human rhinoviruses on optically coated silicon surfaces.

    Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology·2001
    Same author

    Interference-based detection of nucleic acid targets on optically coated silicon.

    Nature biotechnology·2001

    Area of Science:

    • Molecular Biology
    • Genetics
    • Biochemistry

    Background:

    • Plasmid cloning vectors are essential tools in molecular biology.
    • The lacZ gene, encoding beta-galactosidase, is frequently used as a reporter gene.
    • Understanding gene expression regulation is crucial for genetic engineering.

    Purpose of the Study:

    • To characterize beta-galactosidase expression from the pBGP120 plasmid.
    • To investigate the impact of heterologous DNA insertion on beta-galactosidase expression.
    • To determine the regulatory mechanisms controlling lacZ expression from the plasmid.

    Main Methods:

    • Plasmid construction using pBGP120 with EcoRI site near lacZ.
    • Insertion of EcoRI-generated DNA fragments into the plasmid.

    Related Experiment Videos

  • Analysis of beta-galactosidase polypeptide molecular weight and expression levels.
  • Growth in various bacterial strains (cya, lac repressor overproducing) to assess regulation.

    • Main Results:

    • The EcoRI site is within the beta-galactosidase coding sequence.
    • Insertion of DNA fragments in the correct phase resulted in high-level expression of fused proteins.
    • Plasmid-encoded beta-galactosidase showed higher expression than chromosomal, correlating with gene copy number.
    • Expression was dependent on cyclic AMP and regulated by lac repressor.

    Conclusions:

    • Plasmid pBGP120 allows for high-level expression of fused proteins by inserting DNA at the EcoRI site.
    • Gene copy number significantly influences expression levels.
    • Plasmid lacZ expression is subject to both cyclic AMP and lac repressor control.