[29] Transposon mutagenesis for the analysis of protein production, function, and localization
This chapter describes a transposon mutagenesis system that produces multipurpose constructs for the monitoring of protein production, localization, and function. A single mutagenesis generates a large spectrum of alleles, including null, hypomorphic, and conditional alleles, reporter fusions, and e...
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| Veröffentlicht in: | Methods in Enzymology 1999, Vol.303, p.512-532 |
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| container_title | Methods in Enzymology |
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| creator | Ross-Macdonald, Petra Sheehan, Amy Friddle, Carl Roeder, G.Shirleen Snyder, Michael |
| description | This chapter describes a transposon mutagenesis system that produces multipurpose constructs for the monitoring of protein production, localization, and function. A single mutagenesis generates a large spectrum of alleles, including null, hypomorphic, and conditional alleles, reporter fusions, and epitope-insertion alleles. The system, therefore, provides the basis for a wide variety of studies of gene and protein function. The chapter provides comprehensive instructions for use of the new transposons to mutagenize a gene of interest, and for use of the transposon insertion libraries to mutagenize the yeast genome. While the application of these specific transposons is limited to organisms in which the Saccharomyces cerevisiae selectable marker URA3 can be used, the approach is generally applicable to mutagenesis of DNA from any organism for which a transformation and selection system exists. |
| doi_str_mv | 10.1016/S0076-6879(99)03031-1 |
| format | Article |
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| ispartof | Methods in Enzymology, 1999, Vol.303, p.512-532 |
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| source | ScienceDirect eBooks; MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Animals Antibodies Antibodies, Monoclonal Base Sequence beta-Galactosidase - biosynthesis beta-Galactosidase - genetics Cloning, Molecular - methods DNA Primers DNA Transposable Elements Escherichia coli - genetics Genes, Reporter Genetic Vectors Immunoblotting Mice Mutagenesis, Insertional - methods Polymerase Chain Reaction - methods Protein Biosynthesis Proteins - analysis Proteins - genetics Rabbits Recombinant Proteins - analysis Recombinant Proteins - biosynthesis |
| title | [29] Transposon mutagenesis for the analysis of protein production, function, and localization |
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