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

Embryonic stem cell gene targeting using bacteriophage lambda vectors generated by phage-plasmid recombination

T Tsuzuki1, D E Rancourt

  • 1Department of Medical Biochemistry, University of Calgary, Calgary, Alberta T2N 4N1, Canada.

Nucleic Acids Research
|March 21, 1998
PubMed
Summary

This study introduces lambda phage for streamlined gene targeting vector construction, accelerating targeted mutagenesis in molecular medicine. This method simplifies creating specialized mutant animals by bypassing traditional plasmid limitations.

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

WHO 2022 classification of penile and scrotal cancers: updates and evolution.

Histopathology·2022
Same author

Constraints on Cosmic Strings Using Data from the Third Advanced LIGO-Virgo Observing Run.

Physical review letters·2021
Same author

Prospects for observing and localizing gravitational-wave transients with Advanced LIGO, Advanced Virgo and KAGRA.

Living reviews in relativity·2020
Same author

Dataset for the reporting of carcinoma of the bladder-cystectomy, cystoprostatectomy and diverticulectomy specimens: recommendations from the International Collaboration on Cancer Reporting (ICCR).

Virchows Archiv : an international journal of pathology·2020
Same author

Dataset for the reporting of urinary tract carcinoma-biopsy and transurethral resection specimen: recommendations from the International Collaboration on Cancer Reporting (ICCR).

Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc·2019
Same author

Prospects for observing and localizing gravitational-wave transients with Advanced LIGO, Advanced Virgo and KAGRA.

Living reviews in relativity·2018

Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Targeted mutagenesis is crucial for generating animal models in molecular medicine.
  • Constructing targeting vectors (TVs) is a significant bottleneck in gene targeting experiments.
  • Plasmid-based TVs present challenges that can be overcome with alternative methods.

Purpose of the Study:

  • To streamline the generation of targeting vectors (TVs) for targeted mutagenesis.
  • To present a novel method utilizing lambda phage for TV construction.
  • To avoid issues associated with traditional plasmid-based TVs.

Main Methods:

  • Employed a recombination-proficient phage vector, lambda2TK, for TV generation.
  • Utilized conventional restriction-ligation or recombination-mediated methods.

Related Experiment Videos

  • Demonstrated in vivo phage-plasmid recombination for introducing mutations (neo and lacZ-neo).
  • Applied spi selection and size restriction to eliminate single crossover recombinants, ensuring double crossover insertions.

    • Main Results:

    • Successfully generated TVs directly in bacteriophage without plasmid subcloning.
    • Introduced specific mutations into precise genomic locations via double crossover recombination.
    • Achieved efficient generation of phage TVs with double crossover insertions.
    • Reduced the requirement for extensive genomic sequence or restriction site information.

    Conclusions:

    • Lambda phage offers a rapid and efficient alternative for constructing gene targeting vectors.
    • This method simplifies the process of targeted mutagenesis, accelerating the generation of mutant animal models.
    • The direct application of phage vectors bypasses plasmid-related complexities, making gene targeting more accessible.