RECONSTITUTION OF GLUCOSE-UPTAKE AND CHEMOTAXIS IN PSEUDOMONAS-AERUGINOSA GLUCOSE-TRANSPORT DEFECTIVE-MUTANTS

Wild-type glucose uptake and glucose chemotaxis activities were restored in glucose transport defective Pseudomonas aeruginosa strains PFB360 and PFB362 after introduction of plasmid pPZ129, containing a 1.1-kilobase DNA fragment that is essential for the expression of the P. aeruginosa periplasmic glucose binding protein. The restoration of glucose uptake and chemotaxis to wild-type levels in these strains was also achieved by reconstitution with cold-shock fluid and purified glucose binding protein isolated from P. aeruginosa PA01 wild-type strain H103 grown in conditions resulting in the induction of the high-affinity glucose transport system. Glucose uptake was determined by whole cell uptake and shock fluid binding of D-[U-C-14]glucose, using standard filter binding assays. Positive chemotaxis towards glucose was assessed by capillary assays using 10 mM glucose, the amount required for optimal chemotaxis, and judged by plating capillary contents accumulated after 30 min.