Production of recombinant avidin in Escherichia coli.
A recombinant avidin (re-Avd), containing amino acids (aa) 1-123 of the native chicken egg-white Avd, was produced in Escherichia coli. When cells were grown at 37 degrees C production was over 1 microgram/ml, due to altering the codon preference of the first ten codons. The re-Avd was recovered as a soluble protein from cells grown at 25 or 30 degrees C, whereas at 37 degrees C it was mostly insoluble in inclusion bodies. Our results indicated that, despite the potentially harmful biotin-binding activity of Avd, it is possible to produce biologically active Avd in E. coli which then can easily be purified by affinity chromatography on a biotin column in a single step.
Department of Biology, University of Jyväskylä, Finland.
K J
KJ
Airenne
P
P
Sarkkinen
E L
EL
Punnonen
M S
MS
Kulomaa
D000595:Amino Acid Sequence; D001360:Avidin / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon; D017931:DNA Primers; D004926:Escherichia coli / Q000235:genetics* / Q000648:ultrastructure; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012995:Solubility
D003062:Codon; D017931:DNA Primers; D011994:Recombinant Proteins; D001360:Avidin