title: Fast and Efficient Cloning of Cis-Regulatory Sequences for High-Throughput Yeast One-Hybrid Analyses of Transcription Factors.
abstract: Yeast one-hybrid (Y1H) assay has been proven to be a powerful technique to characterize in vivo the interaction between a given transcription factor (TF), or its DNA-binding domain (DBD), and target DNA sequences. Comprehensive characterization of TF/DBD and DNA interactions should allow designing synthetic promoters that would undoubtedly be valuable for biotechnological approaches. Here, we use the ligation-independent cloning system (LIC) in order to enhance the cloning efficiency of DNA motifs into the pHISi Y1H vector. LIC overcomes important limitations of traditional cloning technologies, since any DNA fragment can be cloned into LIC compatible vectors without using restriction endonucleases, ligation, or in vitro recombination.
authors.affiliation: INRA, Institut Jean-Pierre Bourgin, Saclay Plant Sciences, UMR1318, RD10, 78026, Versailles, France.|AgroParisTech, Institut Jean-Pierre Bourgin, Saclay Plant Sciences, UMR1318, RD10, 78026, Versailles, France.
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Kelemen
authors.affiliation: Biochimie et Physiologie Moleculaire des Plantes, UMR 5004, INRA/CNRS/SupAgro-M/UM2, 34060, Montpellier Cedex 1, France.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Przybyla-Toscano
authors.affiliation: Biochimie et Physiologie Moleculaire des Plantes, UMR 5004, INRA/CNRS/SupAgro-M/UM2, 34060, Montpellier Cedex 1, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Tissot
authors.affiliation: INRA, Institut Jean-Pierre Bourgin, Saclay Plant Sciences, UMR1318, RD10, 78026, Versailles, France.|AgroParisTech, Institut Jean-Pierre Bourgin, Saclay Plant Sciences, UMR1318, RD10, 78026, Versailles, France.
authors.forename: Loïc
authors.initials: L
authors.lastname: Lepiniec
authors.affiliation: INRA, Institut Jean-Pierre Bourgin, Saclay Plant Sciences, UMR1318, RD10, 78026, Versailles, France. christian.dubos@supagro.inra.fr.|AgroParisTech, Institut Jean-Pierre Bourgin, Saclay Plant Sciences, UMR1318, RD10, 78026, Versailles, France. christian.dubos@supagro.inra.fr.|Biochimie et Physiologie Moleculaire des Plantes, UMR 5004, INRA/CNRS/SupAgro-M/UM2, 34060, Montpellier Cedex 1, France. christian.dubos@supagro.inra.fr.|Unité de Biochimie et Physiologie Moléculaire des Plantes (B&PMP), 2 Place Pierre Viala, 34060, Montpellier Cedex 02, France. christian.dubos@supagro.inra.fr.
authors.forename: Christian
authors.initials: C
authors.lastname: Dubos
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics*; D020798:Two-Hybrid System Techniques
chemical_list: D014157:Transcription Factors
keywords: Cis-element; Ligase-independent cloning; Transcription factor; Yeast one-hybrid