Development of a Versatile Procedure Based on Natural Transformation for Marker-Free Targeted Genetic Modification in Streptococcus thermophilus
| Autor: | Céline Boutry, Marie Henry de Frahan, Laetitia Fontaine, Philippe Horvath, Brigitte Delplace, Patrick Boyaval, Pascal Hols, Damien Dandoy, Christophe Fremaux |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Genetics
Microbial Genetics Streptococcus thermophilus Mutation Ecology biology Auxotrophy Mutant Mutagenesis (molecular biology technique) Genetics and Molecular Biology biology.organism_classification medicine.disease_cause Applied Microbiology and Biotechnology Phenotype Mutagenesis Insertional Transformation (genetics) Gene Targeting medicine Transformation Bacterial Histidine Sequence Deletion Food Science Biotechnology |
| Zdroj: | Applied and Environmental Microbiology. 76:7870-7877 |
| ISSN: | 1098-5336 0099-2240 |
| Popis: | A versatile natural transformation protocol was established for and successfully applied to 18 of the 19 Streptococcus thermophilus strains tested. The efficiency of the protocol enables the use of in vitro -amplified mutagenesis fragments to perform deletion or insertion of large genetic fragments. Depending on the phenotype linked to the mutation, markerless mutants can be selected either in two steps, i.e., resistance marker insertion and excision using an adapted Cre- loxP system, or in one step using a powerful positive screening procedure as illustrated here for histidine prototrophy. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|