ArticleBMC molecular biology2012
A MultiSite Gateway™ vector set for the functional analysis of genes in the model Saccharomyces cerevisiae.
Article in BMC molecular biology, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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8 citing papers in PubMed, 19 citations in OpenAlex.
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- Strategies for the Development of Industrial Fungal Producing Strains.Journal of fungi (Basel, Switzerland) · 2023Review
- SlKIX8 and SlKIX9 are negative regulators of leaf and fruit growth in tomato.Plant physiology · 2022Article
- The Heat Shock Protein 40-Type Chaperone MASH Supports the Endoplasmic Reticulum-Associated Degradation E3 Ubiquitin Ligase MAKIBISHI1 inFrontiers in plant science · 2021Article
- A user-friendly platform for yeast two-hybrid library screening using next generation sequencing.PloS one · 2018Article
- Quantitative Tandem Affinity Purification, an Effective Tool to Investigate Protein Complex Composition in Plant Hormone Signaling: Strigolactones in the Spotlight.Frontiers in plant science · 2018Article
- The non-JAZ TIFY protein TIFY8 from Arabidopsis thaliana is a transcriptional repressor.PloS one · 2014Article
- A plasmid toolkit for cloning chimeric cDNAs encoding customized fusion proteins into any Gateway destination expression vector.BMC molecular biology · 2013Article
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundRecombinatorial cloning using the Gateway™ technology has been the method of choice for high-throughput omics projects, resulting in the availability of entire ORFeomes in Gateway™ compatible vectors. The MultiSite Gateway™ system allows combining multiple genetic fragments such as promoter, ORF and epitope tag in one single reaction. To date, this technology has not been accessible in the yeast Saccharomyces cerevisiae, one of the most widely used experimental systems in molecular biology, due to the lack of appropriate destination vectors.
resultsHere, we present a set of three-fragment MultiSite Gateway™ destination vectors that have been developed for gene expression in S. cerevisiae and that allow the assembly of any promoter, open reading frame, epitope tag arrangement in combination with any of four auxotrophic markers and three distinct replication mechanisms. As an example of its applicability, we used yeast three-hybrid to provide evidence for the assembly of a ternary complex of plant proteins involved in jasmonate signalling and consisting of the JAZ, NINJA and TOPLESS proteins.
conclusionOur vectors make MultiSite Gateway™ cloning accessible in S. cerevisiae and implement a fast and versatile cloning method for the high-throughput functional analysis of (heterologous) proteins in one of the most widely used model organisms for molecular biology research.
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