Cloning of the netropsin resistance genes from Streptomyces flavopersicus NRRL 2820

✍ Scribed by Tina Stumpp; Sarah Himbert; Josef Altenbuchner

• Publisher: John Wiley and Sons
• Year: 2005
• Tongue: English
• Weight: 122 KB
• Volume: 45
• Category: Article
• ISSN: 0233-111X
• DOI: 10.1002/jobm.200410529

✦ Synopsis

Streptomyces flavopersicus NRRL 2820 (synonym: Streptomyces netropsis DSM40093) is resistant to the N-methylpyrrole-containing oligopeptide antibiotic netropsin. A 9.38 kb DNA-fragment was isolated from a genomic library of Streptomyces flavopersicus using an Escherichia coli-Streptomyces lividans shuttle vector which enables S. lividans to grow on netropsin-containing agar plates. By subcloning, the resistance was conferred to a 5.9 kb Eco RV fragment. DNA sequence analysis of this Eco RV fragment revealed two open reading frames (netP1 , 1556 bp and netP2 , 1773 bp). The deduced proteins share significant similarity to each other (27% identity) and to the large family to ABC-type multidrug resistance proteins. In each protein a conserved transmembrane and ATP binding domain was identified. Deletion analysis showed that both proteins are necessary for netropsin resistance indicating that the proteins form a heterodimeric ABC-transporter exporting netropsin.