Streptomycete host cells of reduced restriction activity.

摘要

Streptomycete host cells having reduced restriction activity are obtained by (a) mutagenising a strain of a Streptomycete species to provide host cells of differing genetic characteristics; (b) growing the surviving host cells and forming protoplasts thereof; (c) transforming the protoplasts with plasmid DNA from another source, such plasmid DNA containing an origin of replication and a selective marker for the Streptomycete host, as well as any DNA sequences necessary for replication and selective culture from the other source; (d) regenerating host cells from the transformed protoplasts containing the plasmid DNA; (e) selecting those host cells which have received the plasmid DNA, and been unable to restrict it, by selecting for expression of the selective marker; (f) confirming that the host cells have reduced restriction activity; and (g) if desired, repeating steps (a) to (f) to reduce further the restriction activity of the host cells and confirm their further reduced restriction activity. Such cells are more able to accept recombinant DNA from another source and are therefore of value for transforming into forms for expression of industrially useful products.