Amplification in Escherichia coli of enzymes involved in genetic recombination: construction of hybrid ColE1 plasmids carrying the structural gene for exonuclease I
| Autor: | N K Alton, Sidney R. Kushner, C L Bassett, Daniel Vapnek |
|---|---|
| Rok vydání: | 1976 |
| Předmět: |
DNA
Bacterial Exonucleases Genotype Mutant DNA Recombinant Extrachromosomal Inheritance Biology medicine.disease_cause Genetic recombination Plasmid Escherichia coli Ultraviolet light medicine Recombination Genetic Genetics Multidisciplinary ColE1 Base Sequence Structural gene Molecular biology Restriction enzyme Genes Genetic Engineering Plasmids Research Article |
| Zdroj: | Proceedings of the National Academy of Sciences. 73:3492-3496 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.73.10.3492 |
| Popis: | Endo-R-HindIII restriction endonuclease fragments obtained from F30 and pMB9 plasmid DNAs were ligated in vitro and used to transform a recB21 recC22 sbcB15 strain of E. coli K-12. The inability of this strain to stably maintain pMB9 alone permitted the isolation of transformants that carried hybrid plasmids containing the sbcB+ allele. These transformants became sensitive to ultraviolet light and recombination defieient and showed a 25-fold increase in the level of exonuclease I activity. The stability of the sbcB hybrid plasmids and their effects on exonuclease I activity have also been determined in wild-type and recA1 genetic backgrounds. The presence of the plasmids results in a 7-fold increase in the level of exonuclease I in a wild-type strain and 15-fold increase in a recA1 strain. The increased activity in the recA1 mutant appears to be a result of increased plasmid stability in this genetic background. |
| Databáze: | OpenAIRE |
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