TARGETED GENE REPLACEMENTS IN ENTERIC BACTERIA USING LINEAR DNA

摘要

The present invention provides a rapid and specific method for targeted gene replacement in bacteria. The modified or disrupted genes or other nucleic acid fragments are transformed into their desired hosts and the transformants are screened for desired integrations using a selectable marker for an altered phenotype. More specifically, the instant invention creates site-directed mutants of enteric bacteria by allelic exchange to identify an altered phenotype that will define the function or effect of the mutated gene. This invention also describes how linear DNA fragments harboring a functional gene can be used for replacement of a mutant gene to restore gene functionality.