The thiostrepton-resistance-encoding gene in Streptomyces laurentii is located within a cluster of ribosomal protein operons.
A common approach to identify and clone biosynthetic gene from an antibiotic-producing streptomycete is to clone the resistance gene for the antibiotic of interest and then use that gene to clone DNA that is linked to it. As a first step toward cloning the genes responsible for the biosynthesis of thiostrepton (Th) in Streptomyces laurentii (Sl), the Th resistance-encoding gene (tsnR) was cloned as a 1.5-kb BamHI-PvuII fragment in Escherichia coli (Ec), and shown to confer Th resistance when introduced into S. lividans TK24. The tsnR-containing DNA fragment was used as a probe to isolate clones from cosmid libraries of DNA in the Ec cosmid vector SuperCos, and pOJ446 (an Ec/streptomycete) cosmid vector. Sequence and genetic analysis of the DNA flanking the tsnR indicates that the Sl tsnR is not closely linked to biosynthetic genes. Instead it is located within a cluster of ribosomal protein operons.
Smith
T M
TM
Department of Medicinal Chemistry BG-20, University of Washington, Seattle 98195, USA.
Jiang
Y F
YF
Shipley
P
P
Floss
H G
HG
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D003360:Cosmids; D004352:Drug Resistance, Microbial / Q000235:genetics; D015723:Gene Library; D005798:Genes, Bacterial; D008780:Methyltransferases / Q000235:genetics*; D008969:Molecular Sequence Data; D005810:Multigene Family; D016254:Mutagenesis, Insertional; D009876:Operon / Q000235:genetics; D010453:Peptide Synthases / Q000235:genetics; D012269:Ribosomal Proteins / Q000235:genetics; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013302:Streptomyces / Q000235:genetics*; D013883:Thiostrepton / Q000494:pharmacology*
D000900:Anti-Bacterial Agents; D012269:Ribosomal Proteins; D008780:Methyltransferases; C035181:rRNA (adenosine-O-2'-)methyltransferase; D010453:Peptide Synthases; D013883:Thiostrepton