UMMS Affiliation

Program in Gene Function and Expression; Department of Biochemistry and Molecular Pharmacology

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Escherichia coli; Gene Deletion; Gene Expression; Genes, Bacterial; Genes, Reporter; *Genetic Markers; Genetic Vectors; Mutagenesis, Insertional; Orotic Acid; Orotidine-5'-Phosphate Decarboxylase; Plasmids; Recombination, Genetic; *Selection, Genetic; Sensitivity and Specificity; Transformation, Bacterial; Two-Hybrid System Techniques


Genetics and Genomics


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.


Biotechniques. 2006 Feb;40(2):179-84. Link to article on publisher's website

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