[Molecular cloning of butirosin biosynthesis genes. I. Construction and characterization of recombinant plasmid DNA No. 2 and No. 11].
B. circulans NRRL-B 3312 is a butirosin-producing strain, butirosin being one of aminoglycoside antibiotics. We used shotgun strategy for molecular cloning of butirosin biosynthesis genes. EcoRI digests of B. circulans NRRL-B 3312 chromosome DNA were ligated to EcoRI digests of plasmid pUB110 with T4-DNA ligase. Recombinant DNA was transformed into competent cells of B. subtilis 168. Kanr transformants were selected on medium containing kanamycin (5 micrograms/ml). Results showed that kanamycin and neomycin resistance levels were higher than that of pUB110 for both No. 2 and No. 11 recombinant plasmid DNA. Restriction enzyme maps were constructed for two recombinant plasmid DNA No. 2 and No. 11 respectively. Southern hybridization showed that there were some homogenous DNA sequences between No. 2 and No. 11 recombinant plasmid DNA and B. circulans NRRL-B 3312 DNA.
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D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D001412:Bacillus subtilis / Q000235:genetics*; D015139:Blotting, Southern; D002076:Butirosin Sulfate / Q000096:biosynthesis*; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D015246:Deoxyribonuclease EcoRI; D010957:Plasmids
D000900:Anti-Bacterial Agents; D004274:DNA, Recombinant; D002076:Butirosin Sulfate; D015246:Deoxyribonuclease EcoRI