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

A simple method for discriminating between cell membrane and cytosolic proteins.

Laura Serna1

  • 1Facultad de Ciencias del Medio Ambiente, Universidad de Castilla-La Mancha, E-45071 Toledo, Spain. laura.serna@uclm.es

The New Phytologist
|February 22, 2005
PubMed
Summary

This study presents a novel method using transgenic plants and hypotonic solutions to easily distinguish between cell membrane and cytosolic green fluorescent protein (GFP)-tagged proteins in plant cells. This technique simplifies protein localization studies.

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

Associations of plasma biomarkers with age in the presenilin-1 E280A autosomal dominant Alzheimer's disease kindred.

The journal of prevention of Alzheimer's disease·2026
Same author

Web-based LAS-FNAME and blood biomarkers in autosomal dominant Alzheimer's disease.

Alzheimer's & dementia (Amsterdam, Netherlands)·2026
Same author

Developing Topics.

Alzheimer's & dementia : the journal of the Alzheimer's Association·2025
Same author

Developing Topics.

Alzheimer's & dementia : the journal of the Alzheimer's Association·2025
Same author

Do grasses have meristemoids?

The New phytologist·2025
Same author

At least two functions for BdMUTE during the development of stomatal complexes in Brachypodium distachyon.

The New phytologist·2025

Area of Science:

  • Plant molecular biology
  • Cell biology
  • Biochemistry

Background:

  • Green fluorescent protein (GFP) fusions are vital for determining protein subcellular localization in transgenic plants.
  • Highly vacuolated plant cells complicate distinguishing between cell wall, cell membrane, and cytosolic GFP-fusion proteins.

Purpose of the Study:

  • To develop a simpler, faster, and more cost-effective method for differentiating cell membrane-bound versus cytosolic GFP-tagged proteins in plant cells.
  • To overcome limitations of traditional methods like plasmolysis tests.

Main Methods:

  • Utilizing transgenic plants expressing GFP-genomic DNA or GFP-cDNA fusions.
  • Observing GFP signal in transgenic protoplasts subjected to hypotonic solutions.
  • Analyzing the diffusion of GFP signal after osmotic vacuole disruption.

Related Experiment Videos

Main Results:

  • Hypotonic treatment of protoplasts allows differentiation between cell membrane and cytosolic localization of GFP-tagged proteins.
  • Soluble GFP-fusion proteins diffuse to the cell's center upon vacuole disruption.
  • Membrane-bound GFP-fusion proteins do not exhibit this diffusion.

Conclusions:

  • The described method offers an easier, faster, and cheaper alternative to subcellular fractionation and immunoelectron microscopy.
  • This technique enhances the ability to accurately determine protein localization in plant cells.