A general in vivo method to amplify the number of copies of a specific gene in one step is described. The method is directly applicable to any selectable gene of Escherichia coli and is based on the Mu-mediated transposition of segments of host chromosomes into the conjugative, multicopy plasmid R6K. Using this method we have cloned the β-hydroxydecanoyl thioester dehydrase structural gene, fabA, into the R6K plasmid. Strains carrying the resultant plasmid produced 13 to 21 times more dehydrase than control strains.
- Molecular cloning
- R6K vector
- in vivo genetic engineering
- β-hydroxydecanoyl thioester dehydrase
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