A Predictive Model of Intein Insertion Site for Use in the Engineering of Molecular Switches
Figure 7
Western blot demonstrating intein splicing at the T134 and S158 sites of XynB.
Intein mutagenized XynB:Tth clones were expressed from lamba phage. Phage infected E.coli were grown on xylanase diagnostic plates as in Fig.6, but at a lower plaque density that allowed scoring individual plaques for xylanase activity. Plaques that displayed distinctively strong blue color, indicative of high xylanase activity, were phagemid rescued to E.coli SOLR cells, and bacterium lysates expressing intein mutagenized XynB:Tth was Western blotted as in Figure 5. Duplicate samples (A and B) of two XynB:Tth clones with mutagenized intein inserted at the T134 and S158 sites are shown at the left. Empty vector control (−) and the intein unmodified XynB is at the right. The T134 and S158 site both support splicing.
