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

Mini-lambda: a tractable system for chromosome and BAC engineering.

Donald L Court1, Srividya Swaminathan, Daiguan Yu

  • 1Molecular Control and Genetics Section, Gene Regulation and Chromosome Biology Laboratory, NCI/FCRDC, National Cancer Institute at Frederick, Building 539/Room 243, P.O. Box B, Frederick, MD 21702, USA. court@ncifcrf.gov

Gene
|October 15, 2003
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

Selective sensitivity of Ph-like B-cell acute lymphoblastic leukemia to BRG1 inhibition identifies a therapeutic vulnerability.

Leukemia·2026
Same author

Accuracy of ophthalmic referral diagnoses by non-ophthalmologists in acute eye care: protocol for a systematic review and meta-analysis.

BMJ open·2026
Same author

RAG-mediated structural variation and its impact on relapse risk in acute lymphoblastic leukemia.

medRxiv : the preprint server for health sciences·2026
Same author

Combining multiplexed assays of variant effect for enhanced BRCA2 variant classification.

Nature communications·2026
Same author

Enhancement of RecET-mediated in vivo linear DNA assembly by a xonA mutation.

PloS one·2026
Same author

Knockdown of the long isoform of the prolactin receptor selectively targets pathogenic immune cells in systemic lupus erythematosus and averts glomerular pathology.

bioRxiv : the preprint server for biology·2026
Same journal

TBX6 promotes proliferation, invasion, and migration in colorectal cancer: Integrated transcriptomic and protein interaction network analysis.

Gene·2026
Same journal

Face/off: phase-specific modeling of lineage plasticity using near-patient models in genitourinary cancers.

Gene·2026
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
See all related articles

A new mini-lambda DNA system enables efficient bacterial artificial chromosome (BAC) engineering using the Red recombination system. This mobile system simplifies in vivo genome engineering and modification of cloned DNA fragments.

Area of Science:

  • Molecular Biology
  • Genetics
  • Microbiology

Background:

  • The bacteriophage lambda (λ) Red recombination system is valuable for engineering large DNA fragments in P1 and bacterial artificial chromosome (BAC) vectors.
  • Current methods require transferring BAC/PAC clones into bacterial cells harboring a defective λ prophage.

Purpose of the Study:

  • To develop a novel, easily transferable mini-lambda DNA system for Red recombination functions.
  • To enable efficient in vivo genome engineering in any *E. coli* strain, including those carrying BACs/PACs.

Main Methods:

  • Generation of a mini-lambda DNA that integrates into the bacterial chromosome as a defective prophage.
  • Electroporation of mini-lambda DNA into *E. coli* strains.
  • Utilizing the mini-lambda system for BAC modification and gene editing.

Related Experiment Videos

Main Results:

  • The mini-lambda DNA provides Red recombination functions and can be introduced into any *E. coli* strain via electroporation.
  • Successful BAC modification by introducing a selectable marker.
  • Creation of a single missense mutation in the human BRCA2 gene within a BAC without a selectable marker.

Conclusions:

  • The mini-lambda system is a simple, mobile tool for in vivo genome engineering via homologous recombination (recombineering).
  • It offers high efficiency for modifying large DNA fragments cloned in BAC/PAC vectors.
  • This system simplifies BAC engineering and gene editing applications.