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Altered Sites® II in vitro
Mutagenesis Systems
By William H. Brondyk
Promega Corporation
Promega's family of Altered Sites® Mutagenesis Systems features positive
antibiotic selection in concert with the widely used approach of oligonucleotide-directed
mutagenesis. The Altered Sites® technology uses oligonucleotides to restore
inactive antibiotic resistance genes. The resulting antibiotic resistance produces
selection for the mutant strand and, thereby, the desired mutation. The Altered Sites®
II Systems use resistance/sensitivity to ampicillin and tetracycline; the Altered Sites®
II Mammalian Systems use resistance/sensitivity to ampicillin and chloramphenicol. By
activating one antibiotic gene (sensitive --> resistant) and simultaneously
inactivating the second antibiotic gene (resistant --> sensitive), many rounds of
mutagenesis are possible.
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