Gene expression after transformation of Streptomyces lividans protoplasts with plasmid pIJ2 DNA.
A new method has been developed for the selection of antibiotic-resistant clones after transformation of Streptomyces protoplasts with plasmid DNA. This method is based on establishing a spatial concentration gradient for the antibiotic, the resistance to which is encoded by the transforming plasmid. By this method, the resistance development of regenerating protoplasts can be followed. The results suggest that antibiotic resistance is inducible. In addition, we were able to show that resident plasmids incompatible with the incoming ones are eliminated when this direct selection principle is used. Moreover, this method, which may facilitate the application of gene technology in Streptomyces, works even though the transformation procedure gives variable results.
D004269:DNA, Bacterial / Q000235:genetics*; D004270:DNA, Circular / Q000235:genetics; D004352:Drug Resistance, Microbial; D019868:Kanamycin Kinase; D009355:Neomycin / Q000494:pharmacology; D010770:Phosphotransferases / Q000235:genetics; D010957:Plasmids; D011523:Protoplasts; D013302:Streptomyces / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D014169:Transformation, Bacterial
D004269:DNA, Bacterial; D004270:DNA, Circular; D010770:Phosphotransferases; D019868:Kanamycin Kinase; D009355:Neomycin