Cloning and expression of a Streptomyces fradiae neomycin resistance gene in Escherichia coli.
A Streptomyces fradiae DNA sequence, which codes for a neomycin phosphotransferase, has been subcloned from the Streptomyces recombinant plasmid pIJ2 [a chimera between the Streptomyces plasmid SLP1.2 and chromosomal DNA containing a neomycin (Nm) resistance gene] into the BamHI restriction enzyme site of pHV14. Three different recombinant plasmids (pWHR1, pWHR2, pWHR3) have been isolated which transform Escherichia coli to Nm resistance. Southern transfer hybridization experiments show that the recombinant plasmids contain the cloned Streptomyces Nm resistance gene, and lysates of E. coli containing the recombinant plasmids were shown to have Nm phosphotransferase activity, demonstrating that a gene from Streptomyces can be expressed in E. coli.
D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004274:DNA, Recombinant / Q000378:metabolism; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D019868:Kanamycin Kinase; D009355:Neomycin / Q000494:pharmacology*; D009693:Nucleic Acid Hybridization; D010770:Phosphotransferases / Q000235:genetics*; D010957:Plasmids; D014176:Protein Biosynthesis; D013302:Streptomyces / Q000235:genetics*
D004274:DNA, Recombinant; D010770:Phosphotransferases; D019868:Kanamycin Kinase; D004262:DNA Restriction Enzymes; D009355:Neomycin