Skip to main content

Journal for Biophysical Chemistry

Biointerphases Cover Image
Figure 4 | Biointerphases

Figure 4

From: Fluorescence-based in situ assay to probe the viability and growth kinetics of surface-adhering and suspended recombinant bacteria

Figure 4

Influence of GFP expression on the adhesion and fluorescence of E. coli K-12 derivative strains. (a) The number of E. coli adhering to bare glass substrates was analyzed for the non-fimbriated empty strain E. coli AAEC191A, strain AAEC191A pHis-GFP that expresses GFP from pHis plasmid upon IPTG induction and strain AAEC191A rpsm-GFP carrying a plasmid to express GFP from the constitutive rpsm promoter. Per strain, bacteria from 20 fields of view (each 200x200 μm) were analyzed. Mean and standard deviation are shown and a two-independent sample two-sided t-test (α = 0.05) was performed. For the E. coli strains tested, the number of adherent bacteria was not significantly different (n.s.) with and without GFP expression. Population variances were not significantly different as tested by a two-sided F-test (α = 0.05). (b) Growth curves of E. coli K-12 derivative strains with and without plasmids for GFP expression. Turbidity of bacterial suspensions in 96 well plates was measured at 600 nm. Mean and standard deviation of a triplicate measurement are shown. (c,d) The fraction of adherent, GFP-fluorescent E. coli was analyzed. The empty strain (AAEC191A, n = 2468) was not fluorescent. GFP expression from both inducible and constitutive promoters yielded similar fractions of fluorescent E. coli (87.3% and 87.2%, respectively). A total of 2852 bacteria carrying the inducible gfp gene (pHis-GFP) and a total of 2036 bacteria carrying the constitutive gfp gene (rpsm-GFP) were analyzed. Shown are means and standard deviations. A two-independent sample two-sided t-test (α = 0.05) was performed and the fraction of fluorescent bacteria was not significantly different (n.s.) for the different GFP expressing E. coli strains. Population variances were not significantly different as tested by a two-sided F-test (α = 0.05).

Back to article page