Systematic mutagenesis of the active site omega loop of TEM-1 beta-lactamase
نویسندگان
چکیده
منابع مشابه
Capturing the mutational landscape of the beta-lactamase TEM-1.
Adaptation proceeds through the selection of mutations. The distribution of mutant fitness effect and the forces shaping this distribution are therefore keys to predict the evolutionary fate of organisms and their constituents such as enzymes. Here, by producing and sequencing a comprehensive collection of 10,000 mutants, we explore the mutational landscape of one enzyme involved in the spread ...
متن کاملPurification of TEM-1 beta-lactamase by immunoaffinity chromatography.
A monoclonal antibody prepared against TEM-1 beta-lactamase was found to compete with penicillins and cephalosporins for binding to the enzyme. The purified antibody preparation was linked to Sepharose 4B and used for immunoaffinity-chromatography purification of TEM-1 beta-lactamase. Elution with either benzylpenicillin or cloxacillin yielded a highly purified, concentrated and active enzyme p...
متن کاملSite-directed mutagenesis using altered beta-lactamase specificity.
Promega's new GeneEditorTM in vitro Site-Directed Mutagenesis System* provides a high efficiency, reliable procedure for the generation and selection of oligonucleotide-directed mutants. The Selection Oligonucleotides provided in this system encode mutations which alter the substrate specificity of beta-lactamase, the gene encoding ampicillin resistance, commonly found on cloning vectors. This ...
متن کاملThe effect of high-frequency random mutagenesis on in vitro protein evolution: a study on TEM-1 beta-lactamase.
For a number of years a major limitation in genetic analysis of protein function has been the inability to introduce multiple substitutions at distant sites that would enable the selection of clusters of mutations required for improved or novel biological functions. In order to achieve this, we have recently developed a novel mutagenesis procedure in which the triphosphate derivatives of a pyri...
متن کاملEvaluation of the Effects of Iron Oxide Nanoparticles on Expression of TEM Type Beta-Lactamase Genes in Pseudomonas Aeruginosa
Pseudomonas aeruginosa is a common cause of surgical-site infections and healthcare-associated infections in the bloodstream, and urinary tract. Iron oxide nanoparticles (IONPs) have shown, to possess antibacterial features. The nanoparticles' status as emerging therapeutic elements has motivated investigators to assess the effects of iron nanoparticles on the expression of TEM type be...
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ژورنال
عنوان ژورنال: Journal of Bacteriology
سال: 1996
ISSN: 0021-9193,1098-5530
DOI: 10.1128/jb.178.7.1821-1828.1996