Current Genetics

, Volume 60, Issue 4, pp 343–350 | Cite as

A modular plasmid system for protein co-localization and bimolecular fluorescence complementation in filamentous fungi

  • Mario Lange
  • Ely Oliveira-Garcia
  • Holger B. Deising
  • Edgar Peiter
Technical Notes

Abstract

To elucidate the function of a protein, it is crucial to know its subcellular location and its interaction partners. Common approaches to resolve those questions rely on the genetic tagging of the gene-of-interest (GOI) with fluorescent reporters. To determine the location of a tagged protein, it may be co-localized with tagged marker proteins. The interaction of two proteins under investigation is often analysed by tagging both with the C- and N-terminal halves of a fluorescent protein. In fungi, the tagged GOI are commonly introduced by serial transformation with plasmids harbouring a single tagged GOI and subsequent selection of suitable strains. In this study, a plasmid system is presented that allows the tagging of several GOI on a single plasmid. This novel double tagging plasmid system (DTPS) allows a much faster and less laborious generation of double-labelled fungal strains when compared with conventional approaches. The DTPS also enables the combination of as many tagged GOI as desired and a simple exchange of existing tags. Furthermore, new tags can be introduced smoothly into the system. In conclusion, the DTPS allows an efficient tagging of GOI with a high degree of flexibility and therefore accelerates functional analysis of proteins in vivo.

Keywords

BiFC Bimolecular fluorescence complementation Co-localization Colletotrichum graminicola Plasmid Protein localization 

Supplementary material

294_2014_429_MOESM1_ESM.pdf (2.2 mb)
Supplementary material 1 (PDF 2242 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Mario Lange
    • 1
  • Ely Oliveira-Garcia
    • 2
  • Holger B. Deising
    • 2
  • Edgar Peiter
    • 1
  1. 1.Plant Nutrition Laboratory, Faculty of Natural Sciences III, Institute of Agricultural and Nutritional SciencesMartin Luther University of Halle-WittenbergHalle (Saale)Germany
  2. 2.Phytopathology and Plant Protection, Faculty of Natural Sciences III, Institute of Agricultural and Nutritional SciencesMartin Luther University of Halle-WittenbergHalle (Saale)Germany

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