Overproduction of theEcoR V endonuclease and methylase
Open Access
- 11 June 1985
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 13 (11), 3823-3839
- https://doi.org/10.1093/nar/13.11.3823
Abstract
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 5-101 of the total cellular proteins, which represents a 50–100 fold increase. A 30 fold overproduction of endonuclease was achieved by randomly positionning 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 attemptsKeywords
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