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

lacZ translation initiation mutations.

L M Munson, G D Stormo, R L Niece

    Journal of Molecular Biology
    |August 25, 1984
    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

    Biotechnology core laboratories: An overview.

    Journal of biomolecular techniques : JBT·2009
    Same author

    Tn5 transposition: a molecular tool for studying protein structure-function.

    Biochemical Society transactions·2006
    Same author

    Identifying muscle regulatory elements and genes in the nematode Caenorhabditis elegans.

    Pacific Symposium on Biocomputing. Pacific Symposium on Biocomputing·2002
    Same author

    A mini-greedy algorithm for faster structural RNA stem-loop search.

    Genome informatics. International Conference on Genome Informatics·2002
    Same author

    Selection of optimal DNA oligos for gene expression arrays.

    Bioinformatics (Oxford, England)·2001
    Same author

    Do mRNAs act as direct sensors of small molecules to control their expression?

    Proceedings of the National Academy of Sciences of the United States of America·2001

    Investigating lacZ gene expression, this study reveals that mRNA secondary structures significantly inhibit protein production. Mutations altering these structures or the Shine-Dalgarno sequence can restore or reduce lacZ expression, impacting translation initiation.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Biochemistry

    Background:

    • The lacZ gene's expression is regulated at the translational level.
    • mRNA secondary structures can mask initiation codons, affecting translation.
    • The Shine-Dalgarno sequence plays a crucial role in ribosome binding and translation initiation.

    Purpose of the Study:

    • To investigate the impact of mRNA secondary structures on lacZ gene expression.
    • To identify mutations that alter mRNA secondary structures and their effect on lacZ expression.
    • To analyze the role of the Shine-Dalgarno sequence in translation initiation under different secondary structure conditions.

    Main Methods:

    • Isolation and characterization of single point mutations in the lacZ gene.
    • Measurement of lacZ expression levels.

    Related Experiment Videos

  • Analysis of mRNA secondary structure formation and its impact on initiation codons.
  • Site-directed mutagenesis to alter Shine-Dalgarno sequence length.

    • Main Results:

    • A stem-and-loop structure in mRNA inhibited lacZ expression 5.8-fold; mutations or deletion relieved this inhibition.
    • Altering the Shine-Dalgarno sequence length had a greater impact on lacZ expression when translation was inhibited by secondary structure.
    • Mutations in the first initiation codon (AUG to GUG) decreased translation initiation tenfold.

    Conclusions:

    • mRNA secondary structures significantly regulate lacZ expression by masking initiation codons.
    • The Shine-Dalgarno sequence's effectiveness is context-dependent, influenced by mRNA secondary structure.
    • Understanding these regulatory mechanisms is crucial for controlling gene expression.