Nucleic Acids Research, 1991, Vol. 19, No. 4 727-731
© 1991
MOLECULAR BIOLOGY |
Gene disruption and gene replacement in Streptomyces via single stranded DNA transformation of integration vectors
Lehrstuhl für Genetik, Fakultät für Biologic, Universität Bielefeld Postfach 8640, D-4800 Bielefeld 1, FRG
* To whom correspondence should be addressed
Received December 13, 1990. Revised January 7, 1991. Accepted January 7, 1991.
For the isolation of single stranded plasmid DNA, various E. coli and E. coli- Streptomvces shuttle plasmids were equipped with the f1 replication origin. The transformation of some representative Streptomyces species with plasmid vectors occurred irrespective of whether single or double stranded DNA was used. In contrast, the transformation of Stmptomyces was 10 to 100 times more efficient when an integration vector was in the single stranded form as opposed to the double stranded form. Streptomyces viridochromogenes was transformed by single stranded DNA integration vectors in order to replace the pat by the tsr gene and generate mutants unable to synthesize phosphinothricin - tripeptide (PTT).
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