title: E. coli BL21(DE3) genomic DNA after OG1RF_10790 overexpression
organism: Escherichia coli BL21(DE3)
description: The OG1RF_10790 gene was amplified using primer set 10790_for_NdeI/10790_rev_BamHI. The 1027 bp fragment was digested with NdeI/BamHI and ligated into pET28a (Novagen), generating pWH51. For overexpression of OG1RF_10790, E. coli BL21DE3 (Novagen) transformed with pWH51 was induced with 1mM IPTG and incubated at 37 degree celsius overnight with shaking at 180 rpm. E. coli BL21DE3 transformed with wild type pET28a was induced under the same conditions as a control. After induction of cultures, 30 ul crude cell lysate was prepared and analyzed by 12% Bis-acrylamide (BioRad) SDS-PAGE and western blot to confirm MTase overexpression (Fig. S1). For western blotting of control pET28a and OG1RF_10790 expression strains, a mouse anti-polyhistidine antibody was used (Cell Signaling).
attributes.tag: strain
attributes.value: BL21(DE3)
attributes.tag: isolation_source
attributes.value: common laboratory strain
attributes.tag: collection_date
attributes.value: 2014-11
attributes.tag: geo_loc_name
attributes.value: USA
attributes.tag: sample_type
attributes.value: genomic DNA
attributes.tag: BioSampleModel
attributes.value: Microbe, viral or environmental