Efficient transformation system for Propionibacterium freudenreichii based on a novel vector
| Autor: | P.H. Pouwels, M.J. van der Werf, R. G. M. Luiten, Johannes Petrus Maria Jore, N. van Luijk |
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| Přispěvatelé: | Centraal Instituut voor Voedingsonderzoek TNO |
| Jazyk: | angličtina |
| Rok vydání: | 2001 |
| Předmět: |
Propionibacterium
Genetic Vectors Molecular Sequence Data Genetics and Molecular Biology Environment medicine.disease_cause Applied Microbiology and Biotechnology Microbiology chemistry.chemical_compound Plasmid medicine Escherichia coli DNA Restriction-Modification Enzymes Replicon Ecology biology Propionibacterium freudenreichii Sequence Analysis DNA biology.organism_classification Molecular biology Transformation (genetics) Restriction enzyme Electroporation chemistry Transformation Bacterial DNA Food Science Biotechnology Plasmids |
| Zdroj: | Applied and Environmental Microbiology, 2, 67, 499-503 |
| Popis: | A 3.6-kb endogenous plasmid was isolated from a Propionibacterium freudenreichii strain and sequenced completely. Based on homologies with plasmids from other bacteria, notably a plasmid from Mycobacterium , a region harboring putative replicative functions was defined. Outside this region two restriction enzyme recognition sites were used for insertion of an Escherichia coli -specific replicon and an erythromycin resistance gene for selection in Propionibacterium . Hybrid vectors obtained in this way replicated in both E. coli and P. freudenreichii . Whereas electroporation of P. freudenreichii with vector DNA isolated from an E. coli transformant yielded 10 to 30 colonies per μg of DNA, use of vector DNA reisolated from a Propionibacterium transformant dramatically increased the efficiency of transformation (≥10 8 colonies per μg of DNA). It could be shown that restriction-modification was responsible for this effect. The high efficiency of the system described here permitted successful transformation of Propionibacterium with DNA ligation mixtures. |
| Databáze: | OpenAIRE |
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