Article Figures & Data
Figures
- FIG. 1.
Comparative analysis of YFP and FbFP expression and in vivo fluorescence in E. coli. E. coli DH5α carrying expression plasmid pRhokHi-2 encoding YFP (A) or FbFP (B) was batch cultured using a BioLector microbioreactor system. The development of biomass (cell density), dissolved oxygen tension (DOT), and FP-mediated fluorescence were permanently online monitored in each well of the MTP. The growth of cultures was monitored with scattered light (I-Io) at an excitation wavelength of 620 nm. Fluorescence emission of FPs was recorded at 540 nm (YFP) and 492 nm (FbFP). Data represent averages from four cultivations (means ± standard deviations). A.U., arbitrary units. Accumulation of YFP (C) and FbFP (D) in E. coli was analyzed at the indicated time points (t1 to t7, marked in panels A and B with black triangles) by immunodetection using specific antisera. All cell extracts were adjusted to the same cell density (corresponding to an OD600 of 10). Accumulation of transcripts of YFP (E) and FbFP (F) was analyzed by RT-PCR using the same cell extracts as for panels C and D. Values are averages from three independent measurements.
- FIG. 2.
Fluorescence characteristics of YFP and FbFP during cell growth. Fluorescence intensities derived from online measurements during E. coli cultivation (Fig. 1A and B) were normalized against the respective cell densities (scattered light intensity I-Io). A.U., arbitrary units.
- FIG. 3.
Comparative expression and in vivo fluorescence of YFP (A) and FbFP (B) in E. coli DH5α cultured in LB medium. Development of biomass (cell density), dissolved oxygen tension (DOT), and FP-mediated autofluorescence were online monitored in each well of the MTP. Background fluorescence from E. coli DH5α (pRhokHi-2) was subtracted. Values are means from four cultivations. (C) Fluorescence characteristics of YFP and FbFP during cell growth in LB medium. Fluorescence intensities derived from online measurements during E. coli cultivation (A and B) were normalized against the respective cell densities (scattered light intensity I-Io). In addition, YFP fluorescence intensity was recalculated (YFPrec) assuming a maturation half time of 1 h. A.U., arbitrary units.
