Isolation and heterologous expression of the Phanerochaete chrysosporium calmodulin gene

We cloned complementary DNA (cDNA) corresponding to protein_id 10767 using transcripts extracted from a culture expressing ligninolytic enzymes, which is registered as a Ca2+-binding protein in the Phanerochaete chrysosporium whole genome database. We used our results to correct an amino acid sequence present in the database. Phylogenetic and southern blot analyses of this re-predicted sequence suggest that this is the sole gene encoding calmodulin (CaM) in this fungus. A P. chrysosporium CaM protein (PcCaM) fused to a His-tag was expressed in Escherichia coli. The His-PcCaM showed appropriate CaM features.

▶ We isolated cDNA of calmodulin (CaM) from Phanerochaete chrysosporium. ▶ The sequence corrected the coding region predicted in whole genome database. ▶ Southern blot analysis suggested the gene is the sole CaM gene in the fungus. ▶ The CaM gene was expressed in Escherichia coli. ▶ The recombinant protein showed appropriate CaM features.