Cloning and analysis of pif, replication and leading regions of the F plasmid |
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| Authors: | Ron Jackson David Cram Animesh Ray Dario DiBerardino and Ron Skurray |
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| Institution: | (1) Department of Microbiology, Monash University, 3168 Clayton, Victoria, Australia;(2) Present address: Department of Molecular Biology, University of Edinburgh, EH9 3JR Edinburgh, Scotland;(3) Present address: Institute of Molecular Biology, University of Oregon, 97403 Eugene, OR, USA |
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| Abstract: | Summary We describe the molecular cloning of BglII fragments of the hybrid plasmid pRS5 (pSC101 and EcoRI fragments of F; f7, f5, f3 and f6). The clones isolated were examined for the expression of F-specified replication, incompatibility, mobilization and inhibition of T7 bacteriophage multiplication. Proteins directed by the BglII clones were labelled in Escherichia coli K12 maxicells and analyzed by SDS-polyacrylamide gel electrophoresis. The sizes of previously reported proteins, encoded by the replication, incompatibility and leading regions encompassed by these plasmids have been confirmed in this study. In addition, the results demonstrate that a pif gene, which encodes an 80,000 dalton polypeptide essential for the inhibition T7 phage multiplication, is located on the BglII fragment that spans the junction of EcoRI fragments f7 and f5. |
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