FIG. 1.
Construction of the pWB980-mini-cbpA1-engB expression plasmid. For B. subtilis expression, we chose a small mini-CbpA, i.e., mini-CbpA1 consisting of a cellulose binding domain, a hydrophilic domain (HLD1), and one cohesin domain (Coh1). Plasmid pWB980, which is a Bacillus expression vector, carries both the constitutively expressed P43 promoter and sacB SP, the engineered levansucrase signal sequence for heterologous protein expression in B. subtilis (23). The sequences coding for the kanamycin resistance marker and levansucrase signal sequence are represented by kmr and sacB SP, respectively. Arrows show the transcription directions for the genes.