Ontology highlight
ABSTRACT:
SUBMITTER: Kuhlman TE
PROVIDER: S-EPMC2847246 | biostudies-literature | 2010 Apr
REPOSITORIES: biostudies-literature
Kuhlman Thomas E TE Cox Edward C EC
Nucleic acids research 20100104 6
We have developed an effective, easy-to-use two-step system for the site-directed insertion of large genetic constructs into arbitrary positions in the Escherichia coli chromosome. The system uses lambda-Red mediated recombineering accompanied by the introduction of double-strand DNA breaks in the chromosome and a donor plasmid bearing the desired insertion fragment. Our method, in contrast to existing recombineering or phage-derived insertion methods, allows for the insertion of very large frag ...[more]