The plasmid CIp10-ECE1 (3 (link), 32 (link)), containing the ECE1 gene and its upstream and downstream intergenic regions, was used as a parental template for site-directed mutagenesis. Site-directed mutagenesis was performed using the QuikChange site-directed mutagenesis system (Agilent). Alanine substitutions in the ECE1 gene were screened by restriction endonuclease digestion, and mutations were confirmed by DNA sequencing. Mutagenized constructs were linearized by digestion with StuI and concentrated by ethanol precipitation prior to transformation.
AlanineDigestionEthanol Gene Intergenic regions Mutations Parental Plasmid Restriction endonuclease Site directed mutagenesis
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Other organizations :
King's College London, Leibniz-Institut für Naturstoff-Forschung und Infektionsbiologie e. V. - Hans-Knöll-Institut (HKI), University of Pittsburgh, University of Aberdeen, Medical Research Council, Universitätsmedizin Göttingen, Friedrich Schiller University Jena
The plasmid CIp10-ECE1 containing the ECE1 gene and its upstream and downstream intergenic regions as a parental template for site-directed mutagenesis
Restriction endonuclease digestion to screen for alanine substitutions in the ECE1 gene
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