Bacteriophage Mu-mediated gene transposition and in vitro cloning of the enterochelin gene cluster of Escherichia coli.
Transposition of chromosomal genes using bacteriophage Mu has been used to obtain a partial order of the nine closely linked genes of the enterochelin-dependent iron transport system of Escherichia coli K-12. Fragments of the ent gene cluster were transposed into the conjugative plasmid RP4 and were characterized by genetic complementation. The partial gene order ( entD, fes), entF, fep, entC, ent(ABEG)...lip was derived using six plasmids which carried overlapping parts of the cluster, and the fep mutations were shown to belong to a single complementation group. Two restriction fragments, one carrying ent(ABCEG) and the other carrying fep, were cloned in vitro using one of the RP4::ent plasmids as a source of DNA enriched in enterochelin system genes. A further restriction fragment, carrying the three remaining genes, entD, fes and entF was cloned directly from the chromosome. The three restriction fragments collectively cover a region of the chromosome 29 kg in length, indicating that the genes of the enterochelin system are clustered but not contiguous.[1]References
- Bacteriophage Mu-mediated gene transposition and in vitro cloning of the enterochelin gene cluster of Escherichia coli. Laird, A.J., Ribbons, D.W., Woodrow, G.C., Young, I.G. Gene (1980) [Pubmed]
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