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 Concept Videos

Overview of Transposition and Recombination02:13

Overview of Transposition and Recombination

Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
DNA-only Transposons02:57

DNA-only Transposons

DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Generation and Single-Cell Transcriptomic Analysis of Hepatocellular Carcinoma Organoids following Drug Treatment.

Journal of visualized experiments : JoVE·2026
Same author

Comparing Multi-Criteria Analysis and Species Distribution Models for Identifying Locust Suitable Habitats in Xinjiang, China.

Biology·2026
Same author

Context-Dependent Effects of Maternal Behaviour on Lamb Growth in Tibetan Sheep.

Animals : an open access journal from MDPI·2026
Same author

Solar radiation may lead to abrupt shifts in centennial-scale temperature variability.

Science bulletin·2026
Same author

Entropy-Driven Spin Transition in Rare Earth Perovskites Enables Feedback Adsorption for Enhanced Acidic Water Oxidation.

Advanced materials (Deerfield Beach, Fla.)·2026
Same author

Global Patterns and Future Dynamics of Four Invasive Cocklebur Species Under Climate Change: Contrasting Climatic and Anthropogenic Drivers.

Biology·2026

Related Experiment Video

Updated: Jul 3, 2026

Generating Transgenic Plants with Single-copy Insertions Using BIBAC-GW Binary Vector
12:08

Generating Transgenic Plants with Single-copy Insertions Using BIBAC-GW Binary Vector

Published on: March 28, 2018

Identifying and genotyping transgene integration loci.

Zhong Liang1, Amy Marie Breman, Brenda R Grimes

  • 1Department of Medical and Molecular Genetics, Indiana University School of Medicine, 975 W Walnut St. IB-130, Indianapolis, IN, 46202, USA. zhliang@iupui.edu

Transgenic Research
|July 10, 2008
PubMed
Summary
This summary is machine-generated.

Identifying transgene insertion sites is crucial for genetic research. This method rapidly verifies integration locations, aiding in breeding strategies and understanding gene expression variations in transgenic mice.

More Related Videos

Bidirectional Retroviral Integration Site PCR Methodology and Quantitative Data Analysis Workflow
12:53

Bidirectional Retroviral Integration Site PCR Methodology and Quantitative Data Analysis Workflow

Published on: June 14, 2017

Analysis of LINE-1 Retrotransposition at the Single Nucleus Level
11:52

Analysis of LINE-1 Retrotransposition at the Single Nucleus Level

Published on: April 23, 2016

Related Experiment Videos

Last Updated: Jul 3, 2026

Generating Transgenic Plants with Single-copy Insertions Using BIBAC-GW Binary Vector
12:08

Generating Transgenic Plants with Single-copy Insertions Using BIBAC-GW Binary Vector

Published on: March 28, 2018

Bidirectional Retroviral Integration Site PCR Methodology and Quantitative Data Analysis Workflow
12:53

Bidirectional Retroviral Integration Site PCR Methodology and Quantitative Data Analysis Workflow

Published on: June 14, 2017

Analysis of LINE-1 Retrotransposition at the Single Nucleus Level
11:52

Analysis of LINE-1 Retrotransposition at the Single Nucleus Level

Published on: April 23, 2016

Area of Science:

  • Genetics
  • Molecular Biology
  • Transgenic Technology

Background:

  • Random germline integration of transgenes is vital for studying gene function.
  • Identifying transgene insertion sites offers practical benefits for breeding and phenotype analysis.

Purpose of the Study:

  • To develop a rapid protocol for identifying and verifying transgene insertion sites in mice.
  • To facilitate genetic marker strategies and distinguish between founder lines.

Main Methods:

  • DNA digestion (NcoI), circularization (T4 DNA ligase), and PCR amplification using transgene-specific primers.
  • DNA sequencing of PCR amplicons and BLASTN search for insertion site identification.
  • Fluorescence in situ hybridization (FISH) analysis for chromosomal localization.

Main Results:

  • A rapid protocol for transgene insertion site identification was successfully developed.
  • FISH analysis confirmed insertion site localization near the end of mouse chromosome 14.
  • The method enables precise identification of transgene integration locations.

Conclusions:

  • This protocol facilitates genotyping for breeding strategies and the construction of mice with multiple genetic markers.
  • Understanding insertion sites is essential for analyzing phenotypes potentially caused by insertional inactivation of endogenous genes.
  • The method aids in distinguishing founder lines with varied transgene expression due to different integration sites.