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Nucleic Acids Research, 1985, Vol. 13, No. 11 3823-3839
© 1985

Articles

Overproduction of the EcoR V endonuclease and methylase

L. Bougueleret1, M.L. Tenchini2, J. Botterman3 and M. Zabeau4

EMBL, Meyerhofstrasse 1, Postfach 102209, D6900 Heidelberg, FRG

Received April 12, 1985. Accepted May 9, 1985.

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 attempts

1Present addresses: Unité d'Informatique Scientifique, Institut Pasteur, 25 me du Dr Roux, 75724 Pans Cedex 15, France

2Present addresses: Departiinento di Biologia e Genetica per le Scienze Mediche, Via Viotti 3, Milano, Italy

3Present addresses: State University of Ghent, Coupure Links, 653 B9000 Ghent

4Present addresses: P.G.S., Plateaustraat 22, B9000 Glient, Belgium


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