Key amino acids residues enhance the ability of CpcR to activate cry gene expression in Bacillus thuringiensis

Typical Bacillus thuringiensis (Bt) produces one or more parasporal crystals composed of insecticidal Cry proteins during the sporulation, and the parasporal crystals and spores are produced from the same cell. Strain Bt LM1212 is different from typical Bt strains in that its crystals and spores are produced in different cells. Previous studies have found that the cell differentiation process of Bt LM1212 is related to the transcription factor CpcR which activates the cry-gene promoters. In addition, CpcR could activate the Bt LM1212 cry35-like gene promoter (P₃₅) when introduced in the heterologous HD73^- strain. It was shown that P₃₅ was only activated in non-sporulating cells. In this study, the peptidic sequences of CpcR homologous proteins found in other strains of the Bacillus cereus group were used as references to identify two key amino acid sites for CpcR activity. The function of these amino acids was investigated by measuring P₃₅ activation by CpcR in strain HD73^-. These results will lay a foundation for the optimization of the insecticidal protein expression system in non-sporulating cells.