A mammalian vector carrying the bleomycin N-acetyltransferase gene from bleomycin-producing Streptomyces verticillus as a selective marker.
A gene, blmB, encodes a bleomycin (Bm) N-acetyltransferase, designated BAT, from Bm-producing Streptomyces verticillus and confers resistance to Bm in Streptomyces and Escherichia coli. COS-1 cells transfected with a plasmid designated pEF-BOS/blmB, in which blmB is under the control of a strong promoter from the human polypeptide chain elongation factor 1alpha, transiently produced BAT. Immuno-cytochemical analysis using an anti-BAT monoclonal antibody revealed that BAT was localized in the nucleus of the blmB-carrying COS-1 cells. NIH/3T3 cells, transfected with pEF-BOS/blmB, stably expressed BAT at least for one month. The stable transformants of blmB showed specific resistance to the Bm family of antibiotics, suggesting that blmB has potential as a selective marker in gene transfer studies with mammalian cells.
Sugiyama
M
M
Institute of Pharmaceutical Sciences, Faculty of Medicine, Hiroshima University, Japan.
Tanaka
M
M
Kumagai
T
T
D000123:Acetyltransferases / Q000235:genetics*; D000617:Aminoglycosides; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins; D001761:Bleomycin / Q000494:pharmacology; D015139:Blotting, Southern; D019556:COS Cells / Q000201:enzymology; D004352:Drug Resistance, Microbial / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005819:Genetic Markers; D005822:Genetic Vectors; D006801:Humans; D020648:Peptide Elongation Factor 1 / Q000187:drug effects / Q000378:metabolism; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics*; D014162:Transfection / Q000235:genetics
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D005819:Genetic Markers; D020648:Peptide Elongation Factor 1; D001761:Bleomycin; C087954:bleomycin N-acetyltransferase; D000123:Acetyltransferases