Overproduction of the EcoR V endonuclease and methylase.
Strains overproducing the EcoR V endonuclease and methylase have been obtained by inserting each of the two genes in expression vectors containing the lambda PL promoter. The methylase is overproduced to a level reaching 5-10% of the total cellular proteins, which represents a 50-100 fold increase. A 30 fold overproduction of endonuclease was achieved by randomly positioning the EndRV gene downstream of the lambda PL promoter. The situation in the endonuclease overproducing clone resembles that encountered in maxi-cells. The strains described here allowed a quick purification of both enzymes in sufficient amounts for crystallisation attempts.
Bougueleret
L
L
Tenchini
M L
ML
Botterman
J
J
Zabeau
M
M
D001483:Base Sequence; D004262:DNA Restriction Enzymes / Q000096:biosynthesis / Q000235:genetics*; D004251:DNA Transposable Elements; D015252:Deoxyribonucleases, Type II Site-Specific; D004790:Enzyme Induction; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D005822:Genetic Vectors; D008780:Methyltransferases / Q000096:biosynthesis / Q000235:genetics*; D010957:Plasmids; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific); D013045:Species Specificity
D004251:DNA Transposable Elements; D008780:Methyltransferases; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific); D004262:DNA Restriction Enzymes; D015252:Deoxyribonucleases, Type II Site-Specific; C027440:GATATC-specific type II deoxyribonucleases