Abstract
The potential of the gfp (green fluorescent protein) gene as a visible marker for tracking hydrogen-producing bacterium of Enterobacter aerogenes was examined. A gfp-and kanamycin-resistance gene-containing vector (pUCGK) was constructed and transformed into E. aerogenes. The fluorescence of the green fluorescent protein (GFP) expression was detected in this fluorescent bacterium under different conditions. Since GFP as a molecular reporter is restricted by its requirement for oxygen in the development of the fluorophore, fluorescence detection for the fluorescent strain grown aerobically and anaerobically was performed, respectively. The oxygen concentration for maturation of the GFP fluorophore in E. aerogenes and the fluorescence detection condition for the fluorescent strain were determined in the hydrogen production system in the present study. By using the GFP technique, cell concentration of E. aerogenes could be easily obtained through the detection of fluorescence intensity. These findings allow the gfp-harboring E. aerogenes to be quantified under an anaerobic condition. As far as we know, we are the first to apply gfp to E. aerogenes for analyzing the hydrogen production process.
