Molecular cloning and in vitro expression of a silent phenoxazinone synthase gene from Streptomyces lividans.
Phenoxazinone synthase (PHS) catalyzes a step in actinomycin D biosynthesis in Streptomyces antibioticus. Two sequences from Streptomyces lividans that hybridize to the phs gene of S. antibioticus have been cloned in Escherichia coli K-12 using the plasmid pBR322. Although there was some similarity in the restriction maps of the two cloned fragments, neither insert appeared to be a direct subset of the other nor of the S. antibioticus phs gene. In vitro expression studies, in a streptomycete coupled transcription-translation system, showed that a 3.98-kb SphI fragment encoded a PHS-related protein. These observations provide additional support for the existence of silent genes for antibiotic production in streptomycetes.
Madu
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AC
Department of Biology, University of Michigan, Ann Arbor 48109.
Jones
G H
GH
D015139:Blotting, Southern; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004587:Electrophoresis, Agar Gel; D015870:Gene Expression; D005798:Genes, Bacterial; D010088:Oxidoreductases / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D015183:Restriction Mapping; D013302:Streptomyces / Q000235:genetics*
D010088:Oxidoreductases; C023944:2-aminophenol oxidase; D004262:DNA Restriction Enzymes