Tightly controlled expression systems for the production and purification of Escherichia coli release factor 1.
The genes for the protein release factors in Escherichia coli have traditionally proven difficult to maintain on high copy plasmids. We have established here systems which provide for both stable maintenance of the release factor 1 gene on such plasmids, as well as high level overproduction of the release factor 1 protein. The gene is maintained under the control of the inducible trc or tac promoters in the presence of very high levels of lac repressor. A simple and rapid scheme for the purification of RF1 from extracts of cultures carrying these plasmids is also described.
Donly
B C
BC
Department of Biochemistry, University of Otago, Dunedin, New Zealand.
Edgar
C D
CD
Williams
J M
JM
Tate
W P
WP
D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins; D005822:Genetic Vectors; D010454:Peptide Termination Factors / Q000096:biosynthesis* / Q000302:isolation & purification; D010957:Plasmids / Q000235:genetics
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D010454:Peptide Termination Factors; C489005:prfA protein, E coli