The use of selection in recovery of transgenic targets for mutation analysis |
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| Institution: | 1. Neurology and PolyTrauma Services, Louis Stokes Cleveland Department of Veterans Affairs Medical Center, Cleveland, OH, USA;2. Department of Neurology, Case Western Reserve University School of Medicine, Cleveland, OH, USA;3. Department of Neurosciences, Case Western Reserve University School of Medicine, Cleveland, OH, USA;1. State Key Laboratory of Urban Water Resource and Environment, School of Environment, Harbin Institute of Technology, Harbin 150090, China;2. School of Environmental Science and Engineering, Southern University of Science and Technology, Shenzhen 518055, China;3. Begbroke Science Park, Department of Engineering Science, University of Oxford, Woodstock Road, Oxford OX5 1PF, United Kingdom;4. Department of Chemical Engineering, National Taiwan University, Taipei 106, Taiwan;5. Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei 106, Taiwan |
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| Abstract: | Transgenic animal mutagenesis assays using lambda shuttle vectors have recently been described for isolation and characterization of spontaneous and chemical induced DNA mutations. Extensive information on lambda and E. coli genetics provides a wealth of techniques to allow selection of mutant target genes. Here we describe the modification of an E. coli host which permits two methods for the direct selection of mutant genes. These methods reduce the number of plates needed to be screened for a comparable amount of frequency data by 20–100-fold and thus provide a significant savings of the materials and time required for the screening of mutations. In addition, mutants selected by these approaches described here may alter or broaden the spectrum of mutations that are recoverable. Ultimately, a combination of selective and nonselective techniques may prove valuable for the analysis of mutations produced in vivo in transgenic animals. |
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