Isolation of gram quantities of EcoRI restriction and modification enzymes from an overproducing strain.

Journal Article (Journal Article)

Structural genes for EcoRI restriction endonuclease and modification methylase have been inserted into the plasmid vector pKC30 (Shimatake, H., and Rosenberg, M. (1981) Nature (Lond.) 292, 128-132) downstream from the bacteriophage lambda pL promoter. Upon induction of pL expression in strains producing a thermolabile lambda cI857 repressor, synthesis of EcoRI polypeptides is enhanced to the extent that after 4 h they represent several per cent of the total cell protein. Purification of activities overproduced in this manner yields preparations of endonuclease and methylase which appear identical to those obtained from conventional sources, with overall yields corresponding to 0.5 to 0.9 g of each enzyme/kg of cell paste.

Full Text

Duke Authors

Cited Authors

  • Cheng, SC; Kim, R; King, K; Kim, SH; Modrich, P

Published Date

  • September 25, 1984

Published In

Volume / Issue

  • 259 / 18

Start / End Page

  • 11571 - 11575

PubMed ID

  • 6088551

International Standard Serial Number (ISSN)

  • 0021-9258


  • eng

Conference Location

  • United States