Expression and purification of plasmid-encoded Thermoplasma acidophilum citrate synthase from Escherichia coli.
The citrate synthase gene from the thermophilic archaebacterium Thermoplasma acidophilum was expressed in Escherichia coli, yielding an active product of the expected molecular weight. Manipulation of the citrate synthase gene in a series of pUC19 constructs showed that the presumed Thermoplasma ribosome binding site is recognized by the E. coli ribosome. A rapid purification of the expression product to homogeneity was achieved, based on the thermostability of Thermoplasma citrate synthase.
Sutherland
K J
KJ
Department of Biochemistry, University of Bath, Claverton Down, UK.
Danson
M J
MJ
Hough
D W
DW
Towner
P
P
D001483:Base Sequence; D001665:Binding Sites; D002950:Citrate (si)-Synthase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D009841:Oligonucleotides; D010957:Plasmids; D013822:Thermoplasma / Q000201:enzymology* / Q000235:genetics
D009841:Oligonucleotides; D002950:Citrate (si)-Synthase