Counter-selectable marker for bacterial-based interaction trap systems
| dc.contributor.author | Meng, Xiangdong | |
| dc.contributor.author | Smith, Robin M. | |
| dc.contributor.author | Giesecke, Astrid V. | |
| dc.contributor.author | Joung, J. Keith | |
| dc.contributor.author | Wolfe, Scot A. | |
| dc.date | 2022-08-11T08:10:15.000 | |
| dc.date.accessioned | 2022-08-23T17:01:00Z | |
| dc.date.available | 2022-08-23T17:01:00Z | |
| dc.date.issued | 2006-03-11 | |
| dc.date.submitted | 2011-04-19 | |
| dc.identifier.citation | Biotechniques. 2006 Feb;40(2):179-84. <a href="http://www.biotechniques.com/BiotechniquesJournal/2006/February/Counter-selectable-marker-for-bacterial-based-interaction-trap-systems/biotechniques-45658.html">Link to article on publisher's website</a> | |
| dc.identifier.issn | 0736-6205 (Linking) | |
| dc.identifier.pmid | 16526407 | |
| dc.identifier.uri | http://hdl.handle.net/20.500.14038/43913 | |
| dc.description.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. | |
| dc.language.iso | en_US | |
| dc.relation | <a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=16526407&dopt=Abstract">Link to Article in PubMed</a> | |
| dc.subject | Escherichia coli | |
| dc.subject | Gene Deletion | |
| dc.subject | Gene Expression | |
| dc.subject | Genes, Bacterial | |
| dc.subject | Genes, Reporter | |
| dc.subject | *Genetic Markers | |
| dc.subject | Genetic Vectors | |
| dc.subject | Mutagenesis, Insertional | |
| dc.subject | Orotic Acid | |
| dc.subject | Orotidine-5'-Phosphate Decarboxylase | |
| dc.subject | Plasmids | |
| dc.subject | Recombination, Genetic | |
| dc.subject | *Selection, Genetic | |
| dc.subject | Sensitivity and Specificity | |
| dc.subject | Transformation, Bacterial | |
| dc.subject | Two-Hybrid System Techniques | |
| dc.subject | Genetics and Genomics | |
| dc.title | Counter-selectable marker for bacterial-based interaction trap systems | |
| dc.type | Journal Article | |
| dc.source.journaltitle | BioTechniques | |
| dc.source.volume | 40 | |
| dc.source.issue | 2 | |
| dc.identifier.legacyfulltext | https://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=1124&context=pgfe_pp&unstamped=1 | |
| dc.identifier.legacycoverpage | https://escholarship.umassmed.edu/pgfe_pp/124 | |
| dc.identifier.contextkey | 1946779 | |
| refterms.dateFOA | 2022-08-23T17:01:00Z | |
| html.description.abstract | <p>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.</p> | |
| dc.identifier.submissionpath | pgfe_pp/124 | |
| dc.contributor.department | Department of Biochemistry and Molecular Pharmacology | |
| dc.contributor.department | Program in Gene Function and Expression | |
| dc.source.pages | 179-84 |
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