title: Counter-selectable marker for bacterial-based interaction trap systems.
abstract: 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.
authors.affiliation: University of Massachusetts Medical School, Worcester, MA 01605, USA.
authors.forename: Xiangdong
authors.initials: X
authors.lastname: Meng
authors.forename: Robin M
authors.initials: RM
authors.lastname: Smith
authors.forename: Astrid V
authors.initials: AV
authors.lastname: Giesecke
authors.forename: J Keith
authors.initials: JK
authors.lastname: Joung
authors.forename: Scot A
authors.initials: SA
authors.lastname: Wolfe
mesh_terms: D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D017353:Gene Deletion; D015870:Gene Expression; D005798:Genes, Bacterial; D017930:Genes, Reporter; D005819:Genetic Markers; D005822:Genetic Vectors; D016254:Mutagenesis, Insertional; D009963:Orotic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D009964:Orotidine-5'-Phosphate Decarboxylase / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D011995:Recombination, Genetic; D012641:Selection, Genetic; D012680:Sensitivity and Specificity; D014169:Transformation, Bacterial; D020798:Two-Hybrid System Techniques
chemical_list: D005819:Genetic Markers; D009963:Orotic Acid; C001242:5-fluoroorotic acid; D009964:Orotidine-5'-Phosphate Decarboxylase