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

Host/vector interactions which affect the viability of recombinant phage lambda clones.

K F Wertman, A R Wyman, D Botstein

    Gene
    |January 1, 1986
    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

    Editorial.

    Molecular biology of the cell·2006
    Same author

    Genomic perspective and cancer.

    Cold Spring Harbor symposia on quantitative biology·2004
    Same author

    The Gene Ontology (GO) database and informatics resource.

    Nucleic acids research·2003
    Same author

    Arrest, adaptation, and recovery following a chromosome double-strand break in Saccharomyces cerevisiae.

    Cold Spring Harbor symposia on quantitative biology·2003
    Same author

    Genome-wide scan of bipolar disorder in 65 pedigrees: supportive evidence for linkage at 8q24, 18q22, 4q32, 2p12, and 13q12.

    Molecular psychiatry·2003
    Same author

    The glycine allele of a glycine/arginine polymorphism in the beta2-adrenergic receptor gene is associated with essential hypertension in a population of Chinese origin.

    American journal of hypertension·2002
    Same journal

    Hierarchical analysis of metabolic phenotype reveals distinct microbiota and circulatory transcriptome in metabolic dysfunction-associated steatotic liver disease.

    Gene·2026
    Same journal

    Mutation T71R enhanced the structural stability and functional activity of wild type superoxide dismutase cloned from soil metagenome.

    Gene·2026
    Same journal

    Reduced ATXN1 expression as an adverse prognostic indicator in Acute myeloid leukemia.

    Gene·2026
    Same journal

    Constructing regulatory networks of Rubisco post-translational modifications: a novel avenue for engineering environment adaptive plants.

    Gene·2026
    Same journal

    Traumatic brain injury enhances fracture healing by upregulating VNN1 to activate the Wnt/β-catenin signaling pathway.

    Gene·2026
    Same journal

    Single-cell transcriptomics reveals CCL2-mediated macrophage-endothelial cell interactions drive apoptosis in varicose veins.

    Gene·2026
    See all related articles

    Recombinant phage lambda clones inhibited on wild-type Escherichia coli can be rescued by introducing the chi sequence or lambda gam+ gene. This research identifies improved host strains for recovering inhibitory genomic sequences.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Microbiology

    Background:

    • Recombinant phage lambda clones derived from human genomic libraries are often inhibited from forming plaques on wild-type Escherichia coli.
    • This inhibition poses a challenge for recovering specific genomic sequences using phage lambda cloning systems.

    Purpose of the Study:

    • To investigate the mechanism underlying the inhibition of specific recombinant phage lambda clones on wild-type E. coli.
    • To identify strategies for overcoming this inhibition and developing improved host strains for phage lambda cloning.

    Main Methods:

    • Recovery of recombinant phage lambda clones from human genomic libraries on defective E. coli strains (recB21 recC22 sbcB15).
    • Testing phage growth on wild-type and various mutant E. coli hosts with altered RecBC enzyme function.

    Related Experiment Videos

  • Introduction of the chi sequence and the lambda gam+ gene into inhibited phage clones.

    • Main Results:

    • Introduction of the chi sequence restored plaque formation on Rec+ cells for one inhibited clone.
    • Insertion of the lambda gam+ gene (encoding an inhibitor of RecBC enzyme) enabled plaque formation on wild-type cells.
    • Host permissiveness correlated with the inactivation of RecBC nucleolytic activities, not recombinational activities.

    Conclusions:

    • The inserted DNA sequences in these phage clones likely limit the production of packageable chromosomes by interacting with host recombination systems.
    • Modified E. coli strains, both recombination-proficient and deficient, were constructed as improved hosts for recovering otherwise inhibitory genomic sequences.
    • Understanding the interplay between phage lambda replication, encapsidation, and host RecBC enzyme function is crucial for optimizing cloning strategies.