Figure 7.

Cloning regions of pSPM-CBD and pSPM-His vectors. The pSPM-CBD (A) and pSPM-His (B) cloning regions harbor the same multiple cloning site (MCS), containing recognition sequences for eight different restriction endonucleases and allowing simple insertion of a target gene, which is fused in-frame to a DNA fragment coding for self-processing module (SPM) of FrpC. MCS-SPM is simultaneously fused in-frame to a chitin-binding domain of Bacillus circulans (CBD) in pSPM-CBD or to a 6× poly-His tag (His) in pSPM-His, respectively. The Asp–Pro bond autocatalytically and specifically cleaved in a fusion protein upon activation by calcium ions is indicated (bold).