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Directed evolution of trimethoprim resistance in Escherichia coli

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Watson, Morgan; Liu, Jian-Wei; Ollis, David

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Directed evolution is a useful tool in the study of enzymes. It is used in this study to investigate the means by which resistance to the antibiotic trimethoprim develops in dihyrofolate reductase from Escherichia coli. Mutants with clinical levels of resistance were obtained after only three generations. After four generations of directed evolution, several mutants were characterized, along with some point mutants made to investigate amino acid changes of interest. Several mutations were found...[Show more]

dc.contributor.authorWatson, Morgan
dc.contributor.authorLiu, Jian-Wei
dc.contributor.authorOllis, David
dc.date.accessioned2015-12-07T22:51:01Z
dc.identifier.issn1742-464X
dc.identifier.urihttp://hdl.handle.net/1885/27268
dc.description.abstractDirected evolution is a useful tool in the study of enzymes. It is used in this study to investigate the means by which resistance to the antibiotic trimethoprim develops in dihyrofolate reductase from Escherichia coli. Mutants with clinical levels of resistance were obtained after only three generations. After four generations of directed evolution, several mutants were characterized, along with some point mutants made to investigate amino acid changes of interest. Several mutations were found to grant resistance to trimethoprim, both by reducing the binding affinity of the enzyme for the drug, and by increasing the activity of the enzyme.
dc.publisherBlackwell Publishing Ltd
dc.sourceThe FEBS Journal
dc.subjectKeywords: dihydrofolate reductase; trimethoprim; antibiotic resistance; article; binding affinity; DNA library; drug protein binding; enzyme activity; enzyme kinetics; Escherichia coli; minimum inhibitory concentration; molecular evolution; nonhuman; point mutation Antibiotic resistance; Dihydrofolate reductase; Directed evolution; Trimethoprim
dc.titleDirected evolution of trimethoprim resistance in Escherichia coli
dc.typeJournal article
local.description.notesImported from ARIES
local.identifier.citationvolume274
dc.date.issued2007
local.identifier.absfor039999 - Chemical Sciences not elsewhere classified
local.identifier.ariespublicationu4005981xPUB50
local.type.statusPublished Version
local.contributor.affiliationWatson, Morgan, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationLiu, Jian-Wei, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationOllis, David, College of Physical and Mathematical Sciences, ANU
local.description.embargo2037-12-31
local.bibliographicCitation.issue10
local.bibliographicCitation.startpage2661
local.bibliographicCitation.lastpage2671
local.identifier.doi10.1111/j.1742-4658.2007.05801.x
dc.date.updated2015-12-07T12:23:42Z
local.identifier.scopusID2-s2.0-34247578707
CollectionsANU Research Publications

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