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

Direct selection for gene replacement events in yeast.

K Struhl

    Gene
    |December 1, 1983
    PubMed
    Summary
    This summary is machine-generated.

    A novel method enables precise gene replacement in Saccharomyces cerevisiae (yeast) at the HIS3 locus. This technique facilitates the direct selection of gene replacement events, simplifying genetic studies in yeast.

    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

    MiR-27b targets PPARγ to inhibit growth, tumor progression and the inflammatory response in neuroblastoma cells.

    Oncogene·2011
    Same author

    Enzymatic labeling of DNA.

    Current protocols in human genetics·2008
    Same author

    Introduction of plasmid DNA into cells.

    Current protocols in neuroscience·2008
    Same author

    Introduction of plasmid DNA into cells.

    Current protocols in protein science·2008
    Same author

    Hybridization analysis of DNA blots.

    Current protocols in protein science·2008
    Same author

    Introduction of plasmid DNA into cells.

    Current protocols in molecular biology·2008

    Area of Science:

    • Molecular Biology
    • Yeast Genetics
    • Gene Editing

    Background:

    • The HIS3 locus in Saccharomyces cerevisiae is crucial for histidine biosynthesis.
    • Efficient gene replacement methods are essential for yeast genetic manipulation and functional studies.

    Purpose of the Study:

    • To develop a robust method for direct gene replacement at the HIS3 locus in yeast.
    • To enable selection of gene replacement events independent of the introduced gene's phenotype.

    Main Methods:

    • Constructed a his3-CYH2 substitution allele by replacing an internal HIS3 region with the CYH2 gene.
    • Transformed a cycloheximide-resistant yeast strain (cyh2r) with the his3-CYH2 allele, creating a cycloheximide-sensitive strain.
    • Introduced new his3 alleles on vectors and selected for cycloheximide resistance to achieve gene replacement.

    Related Experiment Videos

    Main Results:

    • Successfully replaced the wild-type HIS3 locus with the his3-CYH2 substitution.
    • Demonstrated direct selection of gene replacement events by selecting for cycloheximide resistance.
    • The method allows for the introduction of various his3 mutations without direct phenotypic screening.

    Conclusions:

    • The developed method provides a versatile tool for targeted gene replacement in yeast.
    • This approach simplifies the genetic analysis of the HIS3 locus and can be extended to other genomic regions.
    • Facilitates the study of gene function through precise allelic exchange in Saccharomyces cerevisiae.