“Development of modified LUMIER and BiFC assays for the investigation of the putative Varicella Zoster Virus (VZV) nuclear egress complex (NEC) components ORF24 & ORF27”

VIZOSO PINTO MARÍA G; VILLEGAS, J.M; PETER, J; HAASE, R; THOMAE, A; HAAS J; BAIKER, A

Leipzig, Alemania

Congreso; GFV Meeting; 2009

Yeast-two-hybrid analyses have been proven to be an attractive tool for the parallel examination of protein-protein interaction (PPIs). We have perfomed a comprehensive Y2H analysis of all 71 known VZV open reading frames (ORFs). This interactome analysis revelead 173 putatives PPIs within the tested proteins [science 311:239-242]. The Y2H technology, however, is known to exhibit a high rate of false positive and false negative interactions, making the validation of the detected PPIs in different mammalian cell based assay systems extraordinarily important. For the development of such validation assays we focussed on two novel technologies: "LUminescence based Mammalian IntERactome mapping (LUMIER)" [Science 307:1621-1625] and Biomolecular Fluorescence Complementation (BiFC)" [Mol Cell 9:789-798]