METHODS AND MATERIALS FOR THE RAPID AND HIGH VOLUME PRODUCTION OF A GENE KNOCK-OUT LIBRARY IN AN ORGANISM

摘要

The present invention relates to a method for facilitating site directed homologous recombination in an organism to produce mutants comprising: 1) constructing a large insert vector library comprising one or more large insert vectors; 2) constructing a second vector comprising a transposable element; 3) incubating the product of step 1) with the product of step 2) in the presence of a transposase specific for the inverted repeat sequences on the plasmid vector; 4) amplifying the disrupted large insert vector resulting from step 3); 5) introducing the disrupted large insert vector produced by step 4) into a target host cell; and 6) selecting for successful homologous recombination produced by step 5) with the second selectable marker.