Method of selecting recombinant DNA-containing streptomyces
AssigneeEli Lilly and Company
InventorsVirginia A. Birmingham, Eugene T. Seno
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Abstract
A novel method of selecting Streptomyces recombinant DNA-containing host cells and vectors useful in the method are described. The vectors confer tylosin resistance to sensitive Streptomyces host cells and thus provide a convenient method of selecting Streptomyces transformants. The novel tylosin resistance-conferring gene described can be isolated on an .about.2.6 kb KpnI restriction fragment from plasmid pSVB2. Plasmid pSVB2 can be isolated from Streptomyces lividans TK23/pSVB2 (NRRL 15880).
Claims56
A method for selecting a recombinant DNA-containing Streptomyces host cell, said method comprising:
- (a) transforming a tylosin-sensitive, restrictionless Streptomyces host cell with a recombinant DNA cloning vector capable of autonomous replication or integration in said Streptomyces host cell, said vector comprising a DNA sequence that confers resistance to tylosin, and
- (b) culturing said transformed cell under growth conditions suitable for selection for tylosin resistance,
- subject to the limitation that said host cell is susceptible to transformation, cell division and culture.
A recombinant DNA cloning vector capable of autonomous replication or integration in a Streptomyces host cell, said vector comprising a DNA sequence that confers resistance to tylosin in a tylosin-sensitive Streptomyces host cell.
A method for selecting a recombinant DNA-containing Nocardia host cell, said method comprising:
- (a) transforming a tylosin-sensitive, restrictionless Nocardia host cell with a recombinant DNA cloning vector capable of autonomous replication or integration in said Noardia host cell, said vector comprising a DNA sequence that confers resistance to tylosin, and
- (b) culturing said transformed cell under growth conditions suitable for selection for tylosin resistance,
- subject to the limitation that said host cell is susceptible to transformation, cell division and culture.
A Nocardia host cell transformed by a recombinant DNA cloning vector capable of autonomous replication or integration in said Nocardia host cell, said vector comprising a DNA sequence that confers resistance to tylosin in a tylosin-sensitive Nocardia host cell, said Nocardia host cell being tylosin-sensitive when not transformed with said vector.
A recombinant DNA comprising a DNA sequence capable of conferring tylosin resistance on a Streptomyces or Nocardia host cell.
Dependencies are inferred from the claim text.