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

Bacteriophage lambda vector for transducing a cDNA clone library into mammalian cells.

H Okayama, P Berg

    Molecular and Cellular Biology
    |May 1, 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

    Possible involvement of LKB1-AMPK signaling in non-homologous end joining.

    Oncogene·2013
    Same author

    Calcium phosphate mediated gene transfer into established cell lines.

    Methods in molecular biology (Clifton, N.J.)·2011
    Same author

    How different are the indicatrixes of the leaves of various woody plant species?

    Applied optics·2010
    Same author

    Comparisons of indicatrices of desert and urban areas obtained from Landsat MSS data.

    Applied optics·2010
    Same author

    Estimation of soil moisture and components by measuring the degree of spectral polarization with a remote sensing simulator.

    Applied optics·2010
    Same author

    Simulator for remote sensing and its application to soil moisture measurements.

    Applied optics·2010
    Same journal

    Aberrant Expression of miR-25-3p/EZH2 Is Involved in T Cell Activation in Aplastic Anemia.

    Molecular and cellular biology·2026
    Same journal

    Characterization of the m<sup>6</sup>A Epitranscriptome in Fibroblast Senescence.

    Molecular and cellular biology·2026
    Same journal

    Insights into FACT in Cancers with Targeted Therapeutic Implications.

    Molecular and cellular biology·2026
    Same journal

    Human lncRNA, hLinfRNA7 (IDO1-AS) Regulates Cytokine Expression, Tryptophan Catabolism, and Inflammatory Response in Macrophage.

    Molecular and cellular biology·2026
    Same journal

    mTORC1-Dependent Regulation of the CCL24-CCR3 Axis Controls Granuloma Formation and Maintenance in Sarcoidosis.

    Molecular and cellular biology·2026
    Same journal

    The Novel Compound SIC-19 Triggers CUL4B-Mediated Ubiquitination and Degradation of SIK2.

    Molecular and cellular biology·2026
    See all related articles

    Researchers created a bacteriophage lambda NMT vector for efficient mammalian cell transduction using cDNA libraries. This vector enables high-frequency gene transfer and selection of transformed cells, facilitating functional gene discovery.

    Area of Science:

    • Molecular Biology
    • Gene Transfer Technology
    • Mammalian Cell Engineering

    Background:

    • Mammalian cell transduction requires efficient gene delivery systems.
    • Existing methods for introducing cDNA libraries into mammalian cells have limitations.
    • Development of versatile vectors is crucial for functional genomics.

    Purpose of the Study:

    • To develop a novel bacteriophage lambda vector (lambda NMT) for efficient mammalian cell transduction.
    • To enable the introduction of cDNA libraries constructed with the pcD expression vector.
    • To provide a dominant-acting selectable marker for mammalian transformation.

    Main Methods:

    • Construction of the lambda NMT bacteriophage vector.
    • Fusion of a bacterial neo gene to the simian virus 40 early-region promoter for selection.

    Related Experiment Videos

  • Packaging of recombinant phage DNA with pcD-cDNA for in vitro and in vivo propagation.
  • Transfection of mammalian cells and selection of G418-resistant clones.

    • Main Results:

    • The lambda NMT vector efficiently transduced mammalian cells with cDNA libraries.
    • High frequency of G418-resistant transformants (approximately 10^-2) was achieved.
    • Successful transduction and expression of human hypoxanthine-guanine phosphoribosyltransferase (HPRT) in deficient mouse cells.
    • Vector accommodates cDNA inserts up to approximately 9 kilobases.

    Conclusions:

    • The lambda NMT vector is a powerful tool for efficient gene transfer and functional screening in mammalian cells.
    • This system facilitates the identification of functional cDNA clones from libraries.
    • The vector system has significant implications for gene discovery and functional genomics research.