Use of an s-triazine nitrogen source to select for and isolate a recombinant chlorobenzoate-degrading Pseudomonas

Abstract

A novel mating system, which uses the s-triazine nitrogen source (cyanuric acid, CA) as a selective nutritional marker, was developed to evaluate mobility and facilitate identification of chlorobenzoate (CBA) degradation genes in Pseudomonas aeruginosa strain JB2. Matings were done between strain JB2, which is unable to grow with CA, and a CA-utilizer Pseudomonas sp. strain D on 2-CBA-CA and 3-CBA-CA media. Isolates were recovered only from the 2-CBA-CA and were all identified as strain D derivatives. Subtractive hybridization analysis of genomic DNAs from the parental strains and a selected isolate, Pseudomonas sp. strain JPL, was used to determine that the latter organism had acquired approx. 16.1 kb of DNA from strain JB2.