Fig. 4
From: Split intein-mediated selection of cells containing two plasmids using a single antibiotic
SiMPl based on chloramphenicol and ampicillin. a, b Root mean square fluctuation (RMSF) of Cα atoms in chloramphenicol acetyltransferase (PDB ID: 1q23) (a) and TEM-1 β-lactamase (PDB ID: 1zg4) (b) obtained from protein structure fluctuation simulations via the CABS-flex 2.0 web-server20. Flexible regions, within which splice sites were selected, are indicated by black arrows. c, d Crystal structure of TEM-1 β-lactamase (PDB ID: 1zg4) (c) and chloramphenicol acetyltransferase (PDB ID: 1q23) (d). Residues that are part of the active site are represented as gray spheres. Splice sites are represented as sticks and are pointed at by black arrows. Asterisk (*), splice site where the N-terminal fragment of the enzyme showed some independent antibiotic activity. Section sign (§), splice site that did not support bacterial growth. e,h Bar graph showing the transformation efficiency in E. coli TOP10 cells of the indicated plasmids. Values represent mean (± standard error of the mean) of three independent experiments. i Ethidium bromide-stained agarose gel showing plasmid DNA isolated from two randomly picked clones obtained after transformation of E. coli TOP10 cells with the SiMPl plasmids based on chloramphenicol (CAM) and ampicillin (AMP). j Bar graph showing the transformation efficiency in E. coli TOP10 cells of the indicated plasmids. Values represent mean (± standard error of the mean) of three independent experiments. Source data are provided as a Source Data file
