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

Wild-type glucose uptake and glucose chemotaxis activities were restor ed in glucose transport defective Pseudomonas aeruginosa strains PFB36 0 and PFB362 after introduction of plasmid pPZ129, containing a 1.1-ki lobase DNA fragment that is essential for the expression of the P. aer uginosa periplasmic glucose binding protein. The restoration of glucos e 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 g lucose transport system. Glucose uptake was determined by whole cell u ptake and shock fluid binding of D-[U-C-14]glucose, using standard fil ter binding assays. Positive chemotaxis towards glucose was assessed b y capillary assays using 10 mM glucose, the amount required for optima l chemotaxis, and judged by plating capillary contents accumulated aft er 30 min.