Recombinant protein expression at low temperatures under the transcriptional control of the major Escherichia coli cold shock promoter cspA.
A transcriptional gene fusion between the cspA promoter and the lacZ gene was constructed to assess the usefulness of cold shock promoters for low-temperature protein expression. Synthesis of beta-galactosidase was efficiently repressed at 37 degrees C but rapidly induced upon transfer to the 15-to-30 degrees C range, leading to a three- to fivefold increase in specific activity relative to control cultures. Although the initial rates of beta-galactosidase accumulation at 20 degrees C were twice those measured at 15 degrees C, prolonged incubation at 20 degrees C, but not 15 degrees C, led to a dilution of activity due to repression of the promoter and cell division.
Vasina
J A
JA
Department of Chemical Engineering, University of Washington, Seattle 98195, USA.
Baneyx
F
F
D001483:Base Sequence; D003001:Cloning, Molecular; D003080:Cold Temperature; D017931:DNA Primers / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D007700:Kinetics; D007763:Lac Operon; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics*; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D017931:DNA Primers; D004274:DNA, Recombinant; D011994:Recombinant Proteins; D001616:beta-Galactosidase