D. Lyutzkanova et al., A SPECTINOMYCIN RESISTANCE DETERMINANT FROM THE SPECTINOMYCIN PRODUCER STREPTOMYCES-FLAVOPERSICUS, Microbiology, 143, 1997, pp. 2135-2143
The spectinomycin (Sp) resistance determinant from Streptomyces flavop
ersicus was cloned into Streptomyces lividans using the plasmid vector
pIJ699. A plasmid, pDGL15, with a 3.65 kb insert from S. flavopersicu
s conferring resistance to Sp was isolated. DNA sequence analysis of t
he 3651 bp DNA insert revealed four open reading frames (ORFs). The am
ino acid sequence deduced from one ORF (SpcN) showed a high degree of
similarity to an aminoglycoside phosphotransferase (StrN) and from a s
econd one (SpcR) to a regulatory protein (StrR) of the streptomycin bi
osynthesis gene cluster from S. griseus. The two other ORFs were incom
plete and the deduced amino acid sequences showed similarities to an a
midinotransferase encoded in the streptomycin biosynthesis gene cluste
r of S. griseus and to the transposase of IS112, respectively. Express
ion of the spcN gene in E. coli under the control of tac promoter conf
erred Sp resistance to the cells. An enzymic assay confirmed that the
gene product of spcN is an ATP-dependent aminoglycoside phosphotransfe
rase which phosphorylates Sp and actinamine, the aminocyclitol moiety
of Sp.