Properties of hybrid plasmids, consisting of parts of the mini-R1 factorRsc11 andColE1
| Autor: | W. Lindenmaier, W. Boidol, Werner Goebel, G. Siewert, G. Luibrand |
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| Rok vydání: | 1977 |
| Předmět: |
Genetics
ColE1 biology Penicillin Resistance R Factors Extrachromosomal Inheritance EcoRI Colicins Molecular biology SmaI Restriction enzyme Phenotype Plasmid Amp resistance Colicin Escherichia coli biology.protein Hybridization Genetic bacteria Ampicillin Molecular Biology T-DNA Binary system Plasmids |
| Zdroj: | Molecular and General Genetics MGG. 152:231-237 |
| ISSN: | 1432-1874 0026-8925 |
| DOI: | 10.1007/bf00693075 |
| Popis: | In vitro joining of the two small multicopy plasmids Rsc11 and ColE1 by a poly dAdT linker resulted in hybrid plasmids, which determine resistance to ampicillin and immunity to colicin E1. Isolation of the plasmid DNA from single colonies revealed that a variety of hybrid plasmids was formed. Cleavage of these plasmids with restriction endonucleases HinII, HindIII, EcoRI, SmaI and BamI and hybridization with ColE1 demonstrated that they contain different parts of the parent plasmids, Rsc11 and ColE1. Their copy number in the cell is between 6 and 15 per chromosome depending on the plasmid. None of these plasmids can replicate in polA mutants. Replication continues in the presence of chloramphenicol. This suggests that replication can only occur from the ColE1 origin and that the replication function of the Rsc11 part is lost. The hybrid plasmids are compatible with Rsc11 but not with ColE1. The comparison of the physical maps of these Rsc11--ColE1 hybrids with their functions allows a partial determination of the location of ampicillin resistance, replication and incompatibility on the Rsc11 genome. |
| Databáze: | OpenAIRE |
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