Fig. 2
From: Split intein-mediated selection of cells containing two plasmids using a single antibiotic
The SiMPl method based on kanamycin. a, b Schematic showing the main features found on the SiMPl plasmids. pSiMPlk_N (a) and pSiMPlk_C (b) are derivatives of pBAD33 and pTrc99a, respectively. The chloramphenicol resistance gene in pBAD33 is replaced by a fragment of the kanamycin resistance gene encoding amino acids 1 to 118 of aminoglycoside 3′-phosphotransferase followed by the N-terminal fragment of the split gp41-1 intein. Similarly, the ampicillin resistance gene in pTrc99a is replaced by the C-terminal fragment of gp41-1 followed by a fragment of the kanamycin resistance gene encoding amino acids 119 to 271 of aminoglycoside 3′-phosphotransferase. For more efficient splicing, the local exteins (SGY and SSS) are included. c 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. d 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 shown in (a) and (b). e PCR analysis of the SiMPl plasmids isolated from bacteria. pET28a was used as control to show the product obtained after amplification of the full-length kanamycin resistance gene. f Representative fluorescence microscopy images of E. coli TOP10 cells carrying the SiMPl plasmids shown in (a) and (b) induced with 0.1% arabinose and 1 mM IPTG for 3 h. Scale bar, 3 μm. Source data are provided as a Source Data file
