A Predictive Model of Intein Insertion Site for Use in the Engineering of Molecular Switches
Figure 5
Western blot of Tth intein splicing at 20 sites of the XynB xylanase.
A) The Tth intein was inserted into randomly selected 20 sites of the catalytic domain of the XynB generating XynB:Tth. E coli SOLR cell expressing the XynB and intein modified XynB:Tth were lysed, heat treated at 60°C/3 hours and Western blotted. Intein unmodified wild type enzyme XynB (+) and the empty vector control (–) are at the left lanes. Positions of the intein modified precursors (XynB:Tth) and the spliced product (XynB) are marked at the left. B) Insertion sites are mapped out on the crystal structure of XynB. Spice sites are shown as green spheres and non-splice sites are shown as red spheres.
