Abstract: Direct detection of mutagenesis in prokaryotes by reversion of an inactivating mutation (reversion mutation assay), producing a quantitative signal for in vivo mutagenesis, may greatly reduce the amount of test chemicals and labor involved in these assays. Further, transcriptional coupling of ?-lactamase reversion and GFP, translational fusion between ?-lactamase and GFP with stop codon in GFP, and a novel dual reporter to monitor continuous mutagenesis may be used in methods described herein.

Abstract: Direct detection of mutagenesis in prokaryotes by reversion of an inactivating mutation (reversion mutation assay), producing a quantitative signal for in vivo mutagenesis, may greatly reduce the amount of test chemicals and labor involved in these assays. Further, transcriptional coupling of ?-lactamase reversion and GFP, translational fusion between ?-lactamase and GFP with stop codon in GFP, and a novel dual reporter to monitor continuous mutagenesis may be used in methods described herein.

Abstract: Direct detection of mutagenesis in prokaryotes by reversion of an inactivating mutation (reversion mutation assay), producing a quantitative signal for in vivo mutagenesis, may greatly reduce the amount of test chemicals and labor involved in these assays. Further, transcriptional coupling of ?-lactamase reversion and GFP, translational fusion between ?-lactamase and GFP with stop codon in GFP, and a novel dual reporter to monitor continuous mutagenesis may be used in methods described herein.

Abstract: A system for continuous mutagenesis to facilitate directed evolution, the system including DNA polymerases carrying the novel K54E point mutation, and other point mutations including I709N, A759R, D424A (herein called K54E_LF Pol I) and this methods of use to produce and detect lines where mutagenesis is continuous and does not exhibit the usual decline in mutagenesis with sequential cloning.

Abstract: Direct detection of mutagenesis in prokaryotes by reversion of an inactivating mutation (reversion mutation assay), producing a quantitative signal for in vivo mutagenesis, may greatly reduce the amount of test chemicals and labor involved in these assays. Further, transcriptional coupling of ?-lactamase reversion and GFP, translational fusion between ?-lactamase and GFP with stop codon in GFP, and a novel dual reporter to monitor continuous mutagenesis may be used in methods described herein.

Abstract: A system for continuous mutagenesis to facilitate directed evolution, the system including DNA polymerases carrying the novel K54E point mutation, and other point mutations including I709N, A759R, D424A (herein called K54E_LF Pol I) and this methods of use to produce and detect lines where mutagenesis is continuous and does not exhibit the usual decline in mutagenesis with sequential cloning.

Abstract: A reversion mutation assay that is unique in providing a quantitative readout for mutagenesis. This assay is based on the creation of a functional GFP-?-lactamase fusion protein as a reporter providing both antibiotic resistance and fluorescence.

Abstract: A reversion mutation assay that is unique in providing a quantitative readout for mutagenesis. This assay is based on the creation of a functional GFP-?-lactamase fusion protein as a reporter providing both antibiotic resistance and fluorescence. This dual reporter is placed in a multicopy plasmid to increase the number of targets, with a reversion site at the N-terminus. Rare mutations at the reversion site allow read-through of the fusion protein, producing both beta-lactamase (providing antibiotic resistance) and GFP (emitting fluorescence). In the presence of carbenicillin, beta-lactamase production confers a selective advantage that allows amplification of mutant plasmids, raising the level of fluorescence emitted by GFP to levels that are detectable by fluorimetry. A window of time can be found where fluorescence is proportional to the number of mutation events at the reversion site, making fluorescence a quantitative measure of mutagenesis.

Abstract: Super-stable plasmids useful to improve yields of large-scale recombinant gene expression.

Abstract: A reversion mutation assay that is unique in providing a quantitative readout for mutagenesis. This assay is based on the creation of a functional GFP-?-lactamase fusion protein as a reporter providing both antibiotic resistance and fluorescence. This dual reporter is placed in a multicopy plasmid to increase the number of targets, with a reversion site at the N-terminus. Rare mutations at the reversion site allow read-through of the fusion protein, producing both beta-lactamase (providing antibiotic resistance) and GFP (emitting fluorescence). In the presence of carbenicillin, beta-lactamase production confers a selective advantage that allows amplification of mutant plasmids, raising the level of fluorescence emitted by GFP to levels that are detectable by fluorimetry. A window of time can be found where fluorescence is proportional to the number of mutation events at the reversion site, making fluorescence a quantitative measure of mutagenesis.