Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Conservative Site-specific Recombination and Phase Variation

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...

Homologous Recombination

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...

Gene Conversion

Gene Conversion

Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...

In-vitro Mutagenesis

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

Recombinant DNA

Recombinant DNA

Exon Recombination

Exon Recombination

The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes.
Exon shuffling follows “splice frame rules.” Each exon has three reading...

You might also read

Related Articles

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

Sort by

Same author

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

Nature communications·2026

Same author

Rare cancers research: Current state of knowledge and emerging opportunities for prevention and interception.

Journal of the National Cancer Institute·2026

Same author

A single BRCA2 BRC repeat supports viability while multiple repeats ensure resilience under stress.

bioRxiv : the preprint server for biology·2026

Same author

Yes, It Is Worth It: Consider Participant Experiences in the Design of Future Trials.

Cancer prevention research (Philadelphia, Pa.)·2026

Same author

Activity of Direct KRAS(G12C) Inhibitors in Preclinical Models of Pediatric Cancer.

Molecular cancer therapeutics·2025

Same author

5-Hydroxymethylcytosine: a key epigenetic mark in cancer and chemotherapy response.

Epigenetics & chromatin·2025

Same journal

Tracking Synthetic Adhesins on Bacterial Surfaces with Immunofluorescence Microscopy.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Post-Selection Methods for Analyzing mRNA Display Selections and Optimization of Hits.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

High-Performance Computing in Tandem Mass Spectrometry (MS/MS) Peptide Identification.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Engineering and Adapting Disulfide-Containing Proteins to Enable Intracellular Functionality.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

AI-Driven Protein Research: From Prediction to Design.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Methods for the In Vitro Selection of Protein and Peptide Libraries Using mRNA Display.

Methods in molecular biology (Clifton, N.J.)·2026

See all related articles

Search research articles

Related Experiment Video

Updated: May 25, 2026

The Production of C. elegans Transgenes via Recombineering with the galK Selectable Marker

The Production of C. elegans Transgenes via Recombineering with the galK Selectable Marker

Published on: January 11, 2011

Using recombineering to generate point mutations:galK-based positive-negative selection method.

Kajal Biswas¹, Stacey Stauffer, Shyam K Sharan

¹Mouse Cancer Genetics Program, Center for Cancer Research, National Cancer Institute at Frederick, Frederick, MD, USA.

Methods in Molecular Biology (Clifton, N.J.)

|February 14, 2012

Summary

This summary is machine-generated.

This study presents a two-step recombineering method using galactokinase (galK) selection and counterselection for precise point mutation generation in bacterial artificial chromosome (BAC) DNA. This genetic engineering technique efficiently modifies bacterial DNA without restriction enzymes.

More Related Videos

Recombineering Homologous Recombination Constructs in Drosophila

Recombineering Homologous Recombination Constructs in Drosophila

Published on: July 13, 2013

Related Experiment Videos

Last Updated: May 25, 2026

The Production of C. elegans Transgenes via Recombineering with the galK Selectable Marker

The Production of C. elegans Transgenes via Recombineering with the galK Selectable Marker

Published on: January 11, 2011

Recombineering Homologous Recombination Constructs in Drosophila

Recombineering Homologous Recombination Constructs in Drosophila

Published on: July 13, 2013

Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

Recombineering is a powerful, recombination-based genetic engineering technique.
It enables efficient manipulation of bacterial chromosomal and episomal DNA.
Existing methods allow DNA fragment insertion, subcloning, and precise alterations like single nucleotide changes.

Purpose of the Study:

To describe a novel galactokinase (galK)-based two-step method for generating point mutations.
To apply this method to modify bacterial artificial chromosome (BAC) inserts.
To leverage galK selection and counterselection for efficient mutation introduction.

Main Methods:

Utilized recombineering technology for DNA manipulation.
Employed a two-step strategy involving galactokinase (galK).
Implemented selection and counterselection for the presence of galK to introduce point mutations in BAC DNA.

Main Results:

Successfully generated point mutations within BAC inserts.
Demonstrated the efficacy of the galK-based two-step recombineering approach.
Showcased precise DNA alteration capabilities without restriction enzymes.

Conclusions:

The described galK-based two-step recombineering method is highly efficient for generating point mutations in BAC DNA.
This technique offers a precise and versatile tool for genetic engineering in bacteria.
Recombineering provides a powerful alternative to traditional DNA manipulation methods.