Genetic instability in Streptomyces niveus plasmid pSN2: in vivo formation of deletion derivatives.
A plasmid designated pSN2 (molecular size 32.0 kb) was isolated from the wild type of Streptomyces niveus ATCC 19793. To permit phenotypic identification of pSN2 the 1.9 kb BclI fragment was replaced in vitro by the 1.1 kb BclI fragment of pIJ702 carrying the thiostrepton resistance (tsr) gene to form the plasmid pSN3. pSN3 transforms S. lividans to thiostrepton resistance at high frequency and is stably maintained. However, when used to transform S. niveus pSN3 was unstable and produced a 5.5 kb thiostrepton resistant deletion derivative pLG5. pLG5 is also stable and expresses thiostrepton resistance in S. lividans but on transformation of S. niveus was unstable and produced a further thiostrepton resistant derivative, pLG10, of 6.5 kb. pLG5 and pLG10 like pSN3 transform S. lividans at high frequency and produce pocks. DNA hybridizations with a probe derived from pLG5 confirm that pLG5 is derived from DNA sequences present on pSN2 and pSN3.
Hussain
H A
HA
Department of Genetics and Microbiology, Donnan Laboratories, University of Liverpool, UK.
Mitchell
J I
JI
Ritchie
D A
DA
D002872:Chromosome Deletion; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000032:analysis*; D004352:Drug Resistance, Microbial; D004587:Electrophoresis, Agar Gel; D009693:Nucleic Acid Hybridization; D010641:Phenotype; D010957:Plasmids; D011523:Protoplasts; D015183:Restriction Mapping; D013302:Streptomyces / Q000187:drug effects / Q000235:genetics*; D013883:Thiostrepton / Q000494:pharmacology; D014169:Transformation, Bacterial
D004269:DNA, Bacterial; D013883:Thiostrepton