MOLECULAR-CLONING OF A CDNA-ENCODING CALMODULIN FROM NEUROSPORA-CRASSA

A full-length cDNA encoding Neurospora crassa calmodulin was isolated from a lambdaZAP II cDNA expression library. The open reading frame en codes a protein of 148 amino acid residues with a calculated M(r) of 1 6,865 Da. Using site-directed mutagenesis, the complete cDNA was ligat ed into a trc promoter-regulated bacterial expression vector to allow expression of N. crassa calmodulin in E. coli. The expressed protein w as found to be identical to the native protein on the basis of some of its biochemical properties. Finally, Southern analysis of restriction digests of genomic DNA indicates that calmodulin is encoded by a sing le-copy gene.