Clone Of The Enzymes Invovled In Denitrification And Fluorescent Marking Of Pseudomonas Stutzeri LYS-86

The swift growth of industries over the past century has resulted insevere contamination of the environment. Nitrogen is the primarycontamination in water. Biological nitrogen removal is one of the maintechnologies for control of nitrogen pollution.The fluorescence-taggedbacteria is facilitated to the study of its characteristics in biofilm.Pseudomonas stutzeri LYS-86 is an aerobic denitrifying bacteria andhas high activity of nitrogen removal. The strain can be used as a potentialstrain to treat nitrogen-containing wastewater. The genes of nitratereductase NarG, nitrite reductase NirS, nitric oxide reductase NorC andnitrous oxide reductase NosZ were cloned from Pseudomonas stutzeriLYS-86, suggesting the characteristics of denitrification by Pseudomonasstutzeri LYS-86.The strain P. stutzeri LYS-86 had the ability for adsorption to thesurface of alginate calcium gel beads to form biofilm. In order to monitor and investigate the formation and change of biofilm, recombinant plasmidpUT/mini-Tn5-km2-Pnir-gfp which contained nirS promoter wasconstructed. Pnir-gfp was then integrated into the genomic DNA ofP.stutzeri LYS-86 by conjugation. The recombinant strain LYS-Pnir-gfpwas obtained with green fluorescence. And the green fluorescence of thestrain LYS-Pnir-gfp could be kept stably after at least 16 generation. Theother characteristics of P. stutzeri LYS-Pnir-gfp was similar to that of thewild strain of P. stutzeri LYS.