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

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...
Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
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...
Reporter Genes02:11

Reporter Genes

Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
Transposons01:24

Transposons

Transposons, or "jumping genes," are small mobile genetic elements (MGEs) that range from 700 to 40,000 base pairs in length. They are found in all organisms and can move within the same chromosome or transfer to different chromosomes. In some cases, transposons can also jump between different host DNA molecules, such as plasmids or viruses, contributing to genetic variability.Barbara McClintock first discovered these mobile genetic elements in the 1940s while studying maize genetics, and she...
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...

You might also read

Related Articles

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

Sort by
Same author

Rerupture and Total Complication Rate After Single-Incision Power Optimizing Cost-Effective Distal Biceps Repair.

Journal of hand surgery global online·2026
Same author

Integrating Clinical Factors and Parity-Specific Models with Molecular Biomarkers to Better Predict the Risk of Preterm Birth in Asymptomatic Women.

Diagnostics (Basel, Switzerland)·2026
Same author

Incidence of urinary infections and urosepsis after ureteroscopy for stone disease: a systematic review of literature.

Therapeutic advances in urology·2026
Same author

Molecular Dynamics Insights into Ibuprofen Nanocrystal Dissolution Put in the Context of Classical Nucleation Theory.

Molecular pharmaceutics·2026
Same author

International evaluation of the SEIZUre Risk in Encephalitis (SEIZURE) score for predicting acute seizure risk.

BMJ open·2025
Same author

Evaluation of the delivery of the family nurse partnership programme in Scotland during the COVID-19 pandemic.

PloS one·2025

Related Experiment Video

Updated: Jul 8, 2026

Gene Trapping Using Gal4 in Zebrafish
13:34

Gene Trapping Using Gal4 in Zebrafish

Published on: September 29, 2013

Functional fusion proteins by random transposon-based GFP insertion.

Robert Mealer1, Heather Butler, Thomas Hughes

  • 1Department of Cell Biology and Neuroscience, Montana State University, Bozeman, Montana 59717, USA.

Methods in Cell Biology
|December 25, 2007
PubMed
Summary

This study introduces a novel transposon-based method for creating fluorescent sandwich fusion proteins. This technique enables the generation and screening of diverse fusion libraries for biosensor development.

More Related Videos

Imaging Subcellular Structures in the Living Zebrafish Embryo
11:19

Imaging Subcellular Structures in the Living Zebrafish Embryo

Published on: April 2, 2016

Co-expression of Multiple Chimeric Fluorescent Fusion Proteins in an Efficient Way in Plants
09:45

Co-expression of Multiple Chimeric Fluorescent Fusion Proteins in an Efficient Way in Plants

Published on: July 1, 2018

Related Experiment Videos

Last Updated: Jul 8, 2026

Gene Trapping Using Gal4 in Zebrafish
13:34

Gene Trapping Using Gal4 in Zebrafish

Published on: September 29, 2013

Imaging Subcellular Structures in the Living Zebrafish Embryo
11:19

Imaging Subcellular Structures in the Living Zebrafish Embryo

Published on: April 2, 2016

Co-expression of Multiple Chimeric Fluorescent Fusion Proteins in an Efficient Way in Plants
09:45

Co-expression of Multiple Chimeric Fluorescent Fusion Proteins in an Efficient Way in Plants

Published on: July 1, 2018

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Protein Engineering

Background:

  • Fluorescent fusion proteins are crucial tools in molecular biology.
  • Traditional fusion methods involve N- or C-terminal appending, while sandwich fusions offer unique advantages but are challenging to design.
  • Sandwich fusions are valuable for applications like optimizing Förster Resonance Energy Transfer (FRET) biosensors or studying protein conformational changes.

Purpose of the Study:

  • To develop an alternative, efficient protocol for creating functional fluorescent sandwich fusion proteins.
  • To enable the generation of diverse fusion libraries for screening and optimization.
  • To facilitate the complex process of designing novel biosensors.

Main Methods:

  • An in vitro transposon-based random insertion strategy was employed.
  • The coding sequence for a fluorescent protein was randomly inserted into a target protein.
  • This generated a library of sandwich fusion proteins.

Main Results:

  • The protocol successfully created a library of sandwich fusion proteins.
  • The generated library can be screened for desired functional properties.
  • This approach offers a viable solution for complex fusion protein design challenges.

Conclusions:

  • The transposon-based random labeling strategy provides a powerful alternative for constructing fluorescent sandwich fusion proteins.
  • This method simplifies the creation of diverse fusion libraries, aiding in the development of new biosensors.
  • The protocol facilitates the exploration of multiple design solutions for functional fusion proteins.