An Mr 29000 protein is essential for mini-F maintenance in E. coli |
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Authors: | L A Watson S H Phua P L Bergquist H E Lane |
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Institution: | Department of Cell Biology, University of Auckland, Auckland New Zealand |
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Abstract: | Plasmids consisting of mini-F inserted into multicopy vectors were constructed. Derivatives of these hybrid replicons were isolated which contained the transposon Tn5. The polypeptides encoded by these plasmids were identified by Escherichia coli minicell analysis. We show that a previously unidentified polypeptide of 29000 Mr is encoded by the mini-F gene E between 45.1 and 46.2 F kb on the mini-F plasmid map, and that this coding sequence (E gene) is transcribed rightward. Hybrid plasmids carrying Tn5 inserted into the E gene are unable to replicate in a polA- strain. Hence the E protein is essential for mini-F replication. Mutations in the A and B genes of mini-F affect E gene expression, and the results suggest that E protein synthesis is stimulated by A protein. |
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Keywords: | F replication minicells transposon Tn5 restrictionanalysis bp base pairs kb kilobase pairs SDS sodium dodecyl sulfate |
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