Insect larval expression process is optimized by generating fusions with green fluorescent protein

Genomic analysis of high-cell-density recombinantEscherichia coli fermentation and ?cell conditioning? for improved recombinant protein yield

“…The pTH-GFP uv /CAT construct contains a colE1 origin of replication, the trc promoter, the complete gfp uv and CAT genes, a polyhistidine tag, and the ␤-lactamase gene for ampicillin resistance. This construct encodes a unique fusion protein designed to facilitate process monitoring (Cha et al, 1999). W3110 was the strain utilized by Kohara et al (1987) in the creation of the Kohara miniset, and is the strain on the E. coli DNA microarray used in this study.…”

Framework for online optimization of recombinant protein expression in high-cell-densityEscherichia coli cultures using GFP-fusion monitoring

“…For example, online monitoring of green fluorescence protein (GFP) fluorescence to determine recombinant product concentration (Randers-Eichhorn et al, 1997), and online monitoring of firefly luciferase to track intracellular ATP concentration (Lasko and Wang, 1996) have been reported. In the case of GFP, green fluorescence could be monitored online and/or in vivo during fermentation to indicate product level (Albano et al, 1996(Albano et al, , 1998Cha et al, 1997Cha et al, , 1999aDeLisa et al, 1999c;Randers-Eichhorn et al, 1997). Because GFP fluorescence was used to monitor production of a model recombinant protein (CAT) during both low-and high-celldensity cultivations of E. coli (DeLisa et al, 1999c), it was suggested that GFP fluorescence could serve as a sensor for taking a process control action.…”

Expression and purification of recombinant nattokinase in Spodoptera frugiperda cells

“…2A). The decrease in fluorescence intensity after 72 hpi was likely due to proteolysis (Cha et al 1999) as well as cell lysis. The profile of enzyme activity was virtually identical to that of EGFP fluorescence intensity (Fig.…”