A simplified procedure for the rapid identification of recombinant pAT153 plasmids in Escherichia coli HB101 cells.
Insertion of foreign DNA into the unique HindIII site of the high copy number plasmid pAT153 reduces but does not completely abolish the resistance of Escherichia coli HB101 cells to tetracycline. Recombinant DNA-containing colonies could then be phenotypically differentiated from non-recombinant ones by their smaller size on nutrient agar plates with ampicillin and tetracycline at a final concentration of 50 and 4 micrograms/ml, respectively. A wide variety of human cytomegalovirus DNA fragments have been found in pAT153 molecules propagated by the ampicillin-resistant tetracycline-sensitive bacteria selected.
D003587:Cytomegalovirus / Q000235:genetics; D004262:DNA Restriction Enzymes; D015247:Deoxyribonuclease HindIII; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D005822:Genetic Vectors; D010957:Plasmids; D011995:Recombination, Genetic; D014170:Transformation, Genetic
D004262:DNA Restriction Enzymes; D015247:Deoxyribonuclease HindIII