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Nat Commun DOI:10.1038/ncomms9425

Development of potent in vivo mutagenesis plasmids with broad mutational spectra.

Publication TypeJournal Article
Year of Publication2015
AuthorsBadran, AH, Liu, DR
JournalNat Commun
Volume6
Pages8425
Date Published2015 Oct 07
ISSN2041-1723
KeywordsBacteriophage T3, DNA-Directed RNA Polymerases, Drug Resistance, Bacterial, Escherichia coli, Escherichia coli Proteins, Genetic Engineering, Genetic Techniques, Genetic Vectors, High-Throughput Nucleotide Sequencing, Lac Operon, Mutagenesis, Mutation, Plasmids, Promoter Regions, Genetic, Viral Proteins
Abstract

Methods to enhance random mutagenesis in cells offer advantages over in vitro mutagenesis, but current in vivo methods suffer from a lack of control, genomic instability, low efficiency and narrow mutational spectra. Using a mechanism-driven approach, we created a potent, inducible, broad-spectrum and vector-based mutagenesis system in E. coli that enhances mutation 322,000-fold over basal levels, surpassing the mutational efficiency and spectra of widely used in vivo and in vitro methods. We demonstrate that this system can be used to evolve antibiotic resistance in wild-type E. coli in

DOI10.1038/ncomms9425
Pubmed

http://www.ncbi.nlm.nih.gov/pubmed/26443021?dopt=Abstract

Alternate JournalNat Commun
PubMed ID26443021
PubMed Central IDPMC4633624
Grant List / / Howard Hughes Medical Institute / United States
R01 GM065400 / GM / NIGMS NIH HHS / United States