Counter-selectable marker for bacterial-based interaction trap systems
UMass Chan Affiliations
Department of Biochemistry and Molecular PharmacologyProgram in Gene Function and Expression
Document Type
Journal ArticlePublication Date
2006-03-11Keywords
Escherichia coliGene Deletion
Gene Expression
Genes, Bacterial
Genes, Reporter
*Genetic Markers
Genetic Vectors
Mutagenesis, Insertional
Orotic Acid
Orotidine-5'-Phosphate DecarboxylasePlasmidsRecombination, Genetic*Selection, GeneticSensitivity and SpecificityTransformation, BacterialTwo-Hybrid System TechniquesGenetics and Genomics
Metadata
Show full item recordAbstract
Counter-selectable markers can be used in two-hybrid systems to search libraries for a protein or compound that interferes with a macromolecular interaction or to identify macromolecules from a population that cannot mediate a particular interaction. In this report, we describe the adaptation of the yeast URA3/5-FOA counter-selection system for use in bacterial interaction trap experiments. Two different URA3 reporter systems were developed that allow robust counter-selection: (i) a single copy F' episome reporter and (ii) a co-cistronic HIS3-URA3 reporter vector. The HIS3-URA3 reporter can be used for either positive or negative selections in appropriate bacterial strains. These reagents extend the utility of the bacterial two-hybrid system as an alternative to its yeast-based counterpart.Source
Biotechniques. 2006 Feb;40(2):179-84. Link to article on publisher's websitePermanent Link to this Item
http://hdl.handle.net/20.500.14038/43913PubMed ID
16526407Related Resources
Link to Article in PubMedCollections
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