Construction of the glucose isomerase deficient strain of Streptomyces M1033 by homologous recombination

After the establishment of the transformation conditions of Streptomyces di astaticus No.7 Strain M1033, the integration plasmid pXW for homologous rec ombination, which contains a 600 bp fragment of incomplete GI (G138P. G247D ) gene, has been constructed in order to realize the stable overexpression of the GI (G138P. G247D) which is valuable for large-scale industrial produ ction. The Gigene's disruption has been realized by pXW's integration into M1033 chromosomes via homologous recombination and GI deficient strain of S treptomyces M1033 has been obtained. The reliability of introduction of mut ation has been proved by analysis of recombinant fragment and affirmance of existence of the mutation, as well as detection of the stability of the de ficient strain.