IDENTIFICATION OF GENES RESPONSIBLE FOR IN VIVO SURVIVAL OF MICROORGANISMS

摘要

A method for identifying a microorganism having a reduced adaptation to a particular environment comprising the steps of: (I)providing a plurality of microorganisms each of which is independently mutated by the insertional inactivation of a gene with a nucleic acidcomprising a unique marker sequence so that each mutant contains a different marker sequence, or clones of the said microorganism; (2)providing individually a stored sample of each mutant produced by step (1) and providing individually stored nucleic acid comprising theunique marker sequence from each individual mutant; (3) introducing a plurality of mutants produced by step (1) into the said particularenvironment and allowing those microorganisms which are able to do so to grow in the said environment; (4) retrieving microorganismsfrom the said environment or a selected part thereof and isolating the nucleic acid from the retrieved microorganisms; (5) comparing anymarker sequences in the nucleic acid isolated in step (4) to the unique marker sequence of each individual mutant stored as in step (2); and(6) selecting an individual mutant which does not contain any of the marker sequences as isolated in step (4).