Marker reconstitution mutagenesis: a simple and efficient reverse genetic approach
| Autor: | Yun Bao, Mohan K. Balasubramanian, Junqi Huang, Brenda Yuelin Tan, Kavya Bakka, Xie Tang, Anup Padmanabhan, W. Zacheus Cande |
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| Rok vydání: | 2010 |
| Předmět: |
Genetics
Microbial Genetics biology Mutagenesis (molecular biology technique) Bioengineering biology.organism_classification Applied Microbiology and Biotechnology Biochemistry Marker gene Yeast Fungal Proteins Mutagenesis Schizosaccharomyces Schizosaccharomyces pombe Mutant Proteins Exogenous DNA Allele Homologous recombination Gene Biotechnology |
| Zdroj: | Yeast. 28:205-212 |
| ISSN: | 0749-503X |
| DOI: | 10.1002/yea.1831 |
| Popis: | A novel reverse genetic approach termed ‘marker reconstitution mutagenesis’ was designed to generate mutational allelic series in genes of interest. This approach consists of two simple steps which utilize two selective markers. First, using one selective marker, a partial fragment of another selective marker gene is inserted adjacently to a gene of interest by homologous recombination. Second, random mutations are introduced precisely into the gene of interest, together with the reconstitution of the latter selective marker by homologous recombination. This approach was successfully tested for several genes in the fission yeast Schizosaccharomyces pombe. It circumvents the problems encountered with other methods and should be adaptable to any organism that incorporates exogenous DNA by homologous recombination. Copyright © 2010 John Wiley & Sons, Ltd. |
| Databáze: | OpenAIRE |
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