ArticlePloS one2015
A gateway-based system for fast evaluation of protein-protein interactions in bacteria.
Article in PloS one, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed, 11 citations in OpenAlex.
- SoPPIs: a highly parallelized protein-protein-interaction screening method in prokaryotic and eukaryotic hosts.Nucleic acids research · 2026Article
- Development of cell labeling and gene editing tools in urochordateMarine life science & technology · 2025Article
- Bacterial two-hybrid systems evolved: innovations for protein-protein interaction research.Journal of bacteriology · 2025Review
- A Bacterial Two-Hybrid System for In Vivo Assays of Protein-Protein Interactions and Drug Discovery.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Bioengineering ofToxins · 2021Review
- Three Ribosomal Operons ofFrontiers in microbiology · 2021Article
- Article
- pMINERVA: A donor-acceptor system for the in vivo recombineering of scFv into IgG molecules.Journal of immunological methods · 2016Article
- A MultiSite Gateway Toolkit for Rapid Cloning of Vertebrate Expression Constructs with Diverse Research Applications.PloS one · 2016Article
- Self-labelling enzymes as universal tags for fluorescence microscopy, super-resolution microscopy and electron microscopy.Scientific reports · 2015Article
Corrections and comments
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Authors and funding
6 authors at 4 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Protein-protein interactions are important layers of regulation in all kingdoms of life. Identification and characterization of these interactions is one challenging task of the post-genomic era and crucial for understanding of molecular processes within a cell. Several methods have been successfully employed during the past decades to identify protein-protein interactions in bacteria, but most of them include tedious and time-consuming manipulations of DNA. In contrast, the MultiSite Gateway system is a fast tool for transfer of multiple DNA fragments between plasmids enabling simultaneous and site directed cloning of up to four fragments into one construct. Here we developed a new set of Gateway vectors including custom made entry vectors and modular Destination vectors for studying protein-protein interactions via Fluorescence Resonance Energy Transfer (FRET), Bacterial two Hybrid (B2H) and split Gaussia luciferase (Gluc), as well as for fusions with SNAP-tag and HaloTag for dual-color super-resolution microscopy. As proof of principle, we characterized the interaction between the Salmonella effector SipA and its chaperone InvB via split Gluc and B2H approach. The suitability for FRET analysis as well as functionality of fusions with SNAP- and HaloTag could be demonstrated by studying the transient interaction between chemotaxis response regulator CheY and its phosphatase CheZ.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.