Open AccessDirect fluorescence detection of VirE2 secretion by Agrobacterium tumefaciens
Author(s) -
Noga Yaakov,
Yoav Barak,
Idan Pereman,
Peter J. Christie,
Michael Elbaum
Publication year - 2017
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0175273
Subject(s) - agrobacterium tumefaciens , nicotiana tabacum , fluorescence , bimolecular fluorescence complementation , arabidopsis thaliana , green fluorescent protein , alexa fluor , mutant , biology , agrobacterium , in vivo , in vitro , chemistry , plant cell , biophysics , biochemistry , transformation (genetics) , microbiology and biotechnology , genetics , physics , quantum mechanics , gene , yeast
VirE2 is a ssDNA binding protein essential for virulence in Agrobacterium tumefaciens . A tetracysteine mutant (VirE2-TC) was prepared for in vitro and in vivo fluorescence imaging based on the ReAsH reagent. VirE2-TC was found to be biochemically active as it binds both ssDNA and the acidic secretion chaperone VirE1. It was also biologically functional in complementing virE2 null strains transforming Arabidopsis thaliana roots and Nicotiana tabacum leaves. In vitro experiments demonstrated a two-color fluorescent complex using VirE2-TC/ReAsH and Alexa Fluor 488 labeled ssDNA. In vivo , fluorescent VirE2-TC/ReAsH was detected in bacteria and in plant cells at time frames relevant to transformation.