Self cloning in Micromonospora olivasterospora of fms genes for fortimicin A (astromicin) biosynthesis.
We have cloned the seven genes that are responsible for biosynthesis of the antibiotic fortimicin A (FTM A) using a recently developed self-cloning system that employes the plasmid vector pMO116 for Micromonospora olivasterospora. Five chimeric plasmids that restored FTM A production in M. olivasterospora mutants blocked at different biosynthetic steps were isolated by shotgun cloning. Secondary transformation using other non-producing mutants showed that two additional FTM A biosynthetic genes were included on these plasmids, and that at least four of the genes were clustered. Interestingly AN38-1, a non-producing mutant that had a defect in dehydroxylation of a precursor of FTM A, was complemented by the DNA fragment containing a neomycin resistance gene that had been cloned from a neomycin-producing strain (Micromonospora sp. FTM A non-producing strain) in the course of constructing the plasmid vector pMO116. These results clearly show that this novel gene cloning system in Micromonospora is of practical use.
Dairi
T
T
Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., Japan.
Ohta
T
T
Hashimoto
E
E
Hasegawa
M
M
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D005798:Genes, Bacterial; D016236:Genes, fms; D008847:Micromonospora / Q000235:genetics*; D009154:Mutation; D010957:Plasmids; D011523:Protoplasts; D015183:Restriction Mapping; D014170:Transformation, Genetic
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; C023384:fortimicin A