The purification of the Escherichia coli UvrABC incision system.
The UvrA, UvrB and UvrC proteins of Escherichia coli have been purified in good yields to homogeneity with rapid three- or four-step purification procedures. The cloned uvrA and uvrB genes were placed under control of the E. coli bacteriophage lambda PL promoter for amplification of expression. Expression of the uvrC gene could not be amplified by this strategy, however, subcloning of this gene into the replication-defective plasmid pRLM24 led to significant overproduction of the UvrC protein. The purified UvrA protein, with its associated ATPase activity, has a molecular weight of 114,000, the purified UvrB is an 84,000 molecular weight protein and the UvrC protein has a molecular weight of 67,000.
D001426:Bacterial Proteins / Q000302:isolation & purification*; D004249:DNA Damage; D004706:Endodeoxyribonucleases / Q000302:isolation & purification* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D029968:Escherichia coli Proteins; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D010957:Plasmids
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D046911:Macromolecular Substances; D004706:Endodeoxyribonucleases; C036385:endodeoxyribonuclease uvrABC