Site-directed insertion and insertion–deletion mutations in the Escherichia coli chromosome simplified

Abstract

A procedure to produce an exact chromosomal replica of an insertion or insertion–deletion mutation produced in vitro in a plasmid with a ColE 1 origin of replication is presented. This procedure uses a previously described property of recD mutations (Biek DP, Cohen SN. J. Bacteriol. 1986;167:594–603) and is limited by (1) the compatibility of the new mutation with recD; and (2) the presence of some Escherichia coli DNA flanking the mutation.