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

A novel strategy for constructing clustered point mutations.

M Haltiner, T Kempe, R Tjian

    Nucleic Acids Research
    |February 11, 1985
    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

    Getting high to cope with COVID-19: Modelling the associations between cannabis demand, coping motives, and cannabis use and problems.

    Addictive behaviors·2021
    Same author

    Structure of the human Mediator-bound transcription preinitiation complex.

    Science (New York, N.Y.)·2021
    Same author

    An unexpected role of TAFs and TRFs in skeletal muscle differentiation: switching core promoter complexes.

    Cold Spring Harbor symposia on quantitative biology·2008
    Same author

    Transcriptional regulation in Drosophila: the post-genome challenge.

    Functional & integrative genomics·2002
    Same author

    Selectivity of chromatin-remodelling cofactors for ligand-activated transcription.

    Nature·2002
    Same author

    Requirement of tissue-selective TBP-associated factor TAFII105 in ovarian development.

    Science (New York, N.Y.)·2001

    Researchers modified a DNA engineering technique to efficiently create deletions, insertions, and point mutations. This improved method aids in studying DNA regulatory regions like promoters and replication origins.

    Area of Science:

    • Molecular Biology
    • Genetics

    Background:

    • The synthetic linker mutagenesis procedure enables in vitro DNA modification.
    • Previous methods required complex steps for generating mutations.

    Purpose of the Study:

    • To enhance the synthetic linker mutagenesis procedure for easier DNA mutation construction and analysis.
    • To develop novel plasmid vectors for generating nested deletion mutations.
    • To implement a rapid DNA sequencing method for improved efficiency.

    Main Methods:

    • Modification of the synthetic linker mutagenesis protocol.
    • Design of specific plasmid vectors for nested deletion generation.
    • Intramolecular ligation for linker insertion without synthetic DNA.
    • Rapid primer extension sequencing of supercoiled plasmid DNA.

    Related Experiment Videos

    Main Results:

    • Facilitated construction and analysis of deletions, insertions, and clustered point mutations.
    • Enabled insertion of desired linker sequences at deletion endpoints via single intramolecular ligation.
    • Successfully generated nested deletion sets and various mutations in DNA regulatory regions.
    • Demonstrated efficiency in studying transcriptional control regions, including human ribosomal RNA genes.

    Conclusions:

    • The modified protocol offers a rapid and efficient strategy for site-directed mutagenesis.
    • This approach is highly suitable for analyzing DNA regulatory elements.
    • The developed plasmid vectors and sequencing method streamline the study of gene regulation.