title: E. coli BL21(DE3) genomic DNA after OG1RF_11823 overexpression
organism: Escherichia coli BL21(DE3)
description: The OG1RF_11823 gene was amplified using primer set 11823_for_BamHI and 11823_rev_NotI. The 1300 bp fragment was digested with BamHI and NotI and ligated into pGEX-6p-1 (GE Healthcare), generating pWH21. For overexpression of OG1RF_11823, E. coli BL21(DE3) transformed with pWH21 was induced with 1mM IPTG and incubated at 25 degree celsius overnight with shaking at 180 rpm. After induction of cultures, 30 ul crude cell lysate was prepared and analyzed by 12 percent Bis-acrylamide (BioRad) SDS-PAGE and western blot to confirm MTase overexpression (Fig. S1). For western blotting of OG1RF_11823 expression strain, a rabbit anti-GST antibody was used (Thermo Scientific). Genomic DNA was isolated from induced cultures using the protocol described in Supplemental Text.
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