SINGLE CELL LABELING OF A TRICHLOROETHENE-DEGRADING BACTERIUM BY GREEN FLUORESCENT PROTEIN

Abstract

 Phenol-utilizing bacterium, Cupriavidus sp. TW2, is able to degrade Trichloroethene (TCE) by phenol hydroxylase (PH) induced by phenol. Homologous recombination was carried out to insert the tac promoter (P_tac) upstream of PH genes for constitutive expression. The resultant strain, TW2-P, showed the higher TCE degradability without the phenol induction. As a next step, a gene encoding the green fluorescent protein (GPF) was inserted downstream of PH genes by the homologous recombination for single cell monitoring. We succeeded in constructing the strain TW2-PGF that can be monitored as a single cell with a fluorescence microscope. The insertion of GFP gene did not affect the TCE degradability of TW2-P.