Abstract
Pseudomonas fluorescens HK44 represented the first genetically engineered microorganism to be approved in the United States for field release for applications related to subsurface soil bioremediation. In October 1996, strain HK44 was introduced into a replicated semi-contained array of soil lysimeters where its luciferase (luxCDABE)-based bioluminescent response to soil-borne polycyclic aromatic hydrocarbon (PAH) contaminants was detected and monitored for the next two years. At the termination of this experiment, it was decided that the lysimeters remain available for future longer-term monitoring efforts, and were thus covered and left essentially undisturbed until the initiation of a large sampling event in 2010, fourteen years after the original release. Although after extensive sampling culturable HK44 cells were not found, additional molecular and metagenomic analyses indicated that genetic signatures of HK44 cells still persisted, with genes diagnostic for the bioluminescent transposon carried by strain HK44 (luxA and tetA) being found at low concentrations (< 5000 copies/g).
