Fig. 1
From: The iSplit GFP assay detects intracellular recombinant proteins in Bacillus subtilis
Enzymatic activity of GUS(11) and split GFP assay for the GFP11-tagged variants. β-glucuronidase was produced in different amounts by using pBS-Xnt-GUS or pBS-Xnt-GUS11 plasmid series, respectively. Plasmids of the two series harbor the strong constitutive promoter PHpaII and differ in the length of spacer (4–12 nucleotides, indicated as Xnt in the plasmid name) located between the ribosome binding site (Shine-Dalgarno sequence, SD) and gus gene. For the pBS-Xnt-GUS11 plasmid series, a GFP11-tag encoding DNA fragment was fused to gus 3′ end. A Schematic depiction of GUS expression plasmids. Parts of DNA are not drawn to scale; B relative hydrolytic activity of GUS and GUS11; C fluorescence of GUS11 variants determined by split GFP assay in cell lysates of B. subtilis DB430. All measurements were performed in biological and technical triplicates. Error bars indicate the respective standard deviation