Cloning and expression in Streptomyces lividans of a paromomycin phosphotransferase from Streptomyces rimosus Forma paromomycinus.
The paromomycin producing organism Streptomyces rimosus forma paromomycinus is resistant to this antibiotic and contains a phosphotransferase which inactivates paromomycin. The gene encoding this enzyme has been inserted in the Streptomyces vector pIJ702 and then cloned in Streptomyces lividans, selecting for paromomycin-resistance. Three plasmids have been isolated and one of them, pMJ1, contains a 2.2 kb insert with a single HindIII restriction site. Insertion of foreign DNA in this site blocks the expression of the phosphotransferase enzyme indicating that it is within the cloned gene. These findings provide a new dominant selective marker for Streptomyces cloning vectors with the versatility of insertional inactivation.
D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes / Q000378:metabolism; D015247:Deoxyribonuclease HindIII; D010770:Phosphotransferases / Q000096:biosynthesis / Q000235:genetics*; D017853:Phosphotransferases (Alcohol Group Acceptor); D010957:Plasmids; D013302:Streptomyces / Q000235:genetics*
D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C043295:paromomycin phosphotransferase; D004262:DNA Restriction Enzymes; D015247:Deoxyribonuclease HindIII