Localizing protein-protein interactions by bimolecular fluorescence complementation in planta

Vitaly Citovsky, Yedidya Gafni, Tzvi Tzfira

Research output: Contribution to journalArticlepeer-review

45 Scopus citations

Abstract

The application of novel assays for basic cell research is tightly linked to the development of easy-to-use and versatile tools and protocols for implementing such technologies for a wide range of applications and model species. The bimolecular fluorescence complementation (BiFC) assay is one such novel method for which tools and protocols for its application in plant cell research are still being developed. BiFC is a powerful tool which enables not only detection, but also visualization and subcellular localization of protein-protein interactions in living cells. Here we describe the application of BiFC in plant cells while focusing on the use of our versatile set of vectors which were specifically designed to facilitate the transformation, expression and imaging of protein-protein interactions in various plant species. We discuss the considerations of using our system in various plant model systems, the use of single versus multiple expression cassettes, the application of our vectors using various transformation methods and the use of internal fluorescent markers which can assist in signal localization and easy data acquisition in living cells.

Original languageEnglish
Pages (from-to)196-206
Number of pages11
JournalMethods
Volume45
Issue number3
DOIs
StatePublished - Jul 2008
Externally publishedYes

Bibliographical note

Funding Information:
The work in our laboratories is supported by grants from NIH, NSF, USDA, BARD, BSF and CDR-USAID to VC, from ISF and BARD to YG and from CPBR, BARD and BRDC to TT.

Funding

The work in our laboratories is supported by grants from NIH, NSF, USDA, BARD, BSF and CDR-USAID to VC, from ISF and BARD to YG and from CPBR, BARD and BRDC to TT.

FundersFunder number
BRDC
CDR-USAID
National Science Foundation
National Institutes of Health
U.S. Department of Agriculture
Consortium for Plant Biotechnology Research
BARD
United States-Israel Binational Science Foundation
Israel Science Foundation

    Keywords

    • Confocal
    • Fluorescent proteins
    • Imaging
    • Plants
    • Protein interactions
    • Transformation
    • Vector system
    • pSAT

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