Figure 4
From: Environmental and intracellular regulation of Francisella tularensis ripA
Tetracysteine tag construction and expression. (a) Graphical depiction of F. tularensis LVS ripA locus showing the location of SOE PCR primers used to insert the C terminal TC tag (marked in gray). (b) Nucleotide and amino acid sequence of the C terminal TCtag showing the overlapping sequence of the SOE PCR primers. (c) In gel fluorescence of RipA-TC (black arrow) from dilution series of F. tularensis LVS (plasmid) pKK ripA'-TC and F. tularensis LVS (chromosomal)ripA'-TC using 6000 ng to 60 ng total protein of whole cell lysates. F. tularensis LVS lysates (wt) used as a non TC tagged control displaying three non specific bands (gray arrows) at a higher molecular weight than RipA-TC.