Strategy for deletion of complete open reading frames in Saccharomyces cerevisiae
| Autor: | Ines Eberhardt, Stefan Hohmann |
|---|---|
| Rok vydání: | 1995 |
| Předmět: |
Genetics
Models Genetic biology Saccharomyces cerevisiae General Medicine biology.organism_classification Polymerase Chain Reaction law.invention Open Reading Frames Restriction site Restriction enzyme Open reading frame chemistry.chemical_compound Transformation Genetic chemistry law biology.protein Gene Gene Deletion Polymerase chain reaction Polymerase DNA Plasmids |
| Zdroj: | Current Genetics. 27:306-308 |
| ISSN: | 1432-0983 0172-8083 |
| DOI: | 10.1007/bf00352097 |
| Popis: | The classical disruption method for yeast genes is by using in vitro deletion of the gene of interest, or of a part of it, with restriction enzymes. We are now routinely using a strategy that takes advantage of polymerase chain reactions (PCRs) which amplify large pieces of DNA. Since this approach results in a complete, precise deletion of the open reading frame, which is replaced by a unique restriction site, the ligated PCR can be used for the insertion of different markers or for two-step gene disruptions without an inserted marker. As we have now used this strategy for the deletion of more than ten genes we have in this report included some hints based on our experience. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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