Patent ID: 6140129
Filing Date: 2000-10-31
Classification: C12N

Abstract:
A method of introducing exogenous cloned DNA into a bacterial chromosome in which the transposon Tn5 and the FLP recombinase are functional in vivo, comprising the steps of:(a) introducing FLP recombination target sites (FRTs) permanently at random locations in a bacterial chromosome using a plasmid vector that contains an FRT within a modified transposon, two selectable markers, and a removable replication origin;(b) mapping the introduced FRT;(c) cloning exogenous DNA into a plasmid vector comprising two FRT sites, two selectable markers, and a removable replication origin;(d) removing the replication origin in the vector of step (c);(e) introducing the altered plasmid vector of step (d) into bacterial cells, wherein the bacteria cells comprise a functional FLP recombinase; and(f) obtaining targeted integrants.