Uses of beta-galactosidase tag in on-line monitoring production of fusion proteins and gene expression in Escherichia coli.
A simple method for monitoring and quantifying automatically the production by fermentation of beta-galactosidase fusion proteins, making use of the remaining activity of the beta-galactosidase part, is considered. A hybrid protein carrying the major antigenic domain of foot-and-mouth disease virus C1 joined at the N-terminus of beta-galactosidase has been expressed in Escherichia coli. The yield of the chimeric protein has been monitored by flow injection analysis (FIA) during batch fermentations at laboratory scale, and a high correlation between values of product concentration from FIA and from immunological quantizations has been obtained. Because of the possibility of employing FIA in large-scale experiments, and the high sampling frequency, versatility, and reproducibility offered by this method, we propose FIA as a general, simple, quick, flexible, and reliable instrument for both monitoring the yield of recombinant proteins produced industrially, and performing basic research at laboratory scale.
Benito
A
A
Institut de Biologia Fonamental, Universitat Autònoma de Barcelona, Spain.
Valero
F
F
Lafuente
J
J
Vidal
M
M
Cairo
J
J
Solà
C
C
Villaverde
A
A
D005537:Aphthovirus / Q000235:genetics*; D001322:Autoanalysis / Q000295:instrumentation / Q000379:methods; D015153:Blotting, Western / Q000379:methods; D002213:Capsid / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D036022:Capsid Proteins; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics*; D005285:Fermentation; D015870:Gene Expression; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D001616:beta-Galactosidase / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification
D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase