Molecular cloning of a cDNA encoding calmodulin from Neurospora crassa.
A full-length cDNA encoding Neurospora crassa calmodulin was isolated from a lambda ZAP II cDNA expression library. The open reading frame encodes a protein of 148 amino acid residues with a calculated M(r) of 16,865 Da. Using site-directed mutagenesis, the complete cDNA was ligated into a trc promoter-regulated bacterial expression vector to allow expression of N. crassa calmodulin in E. coli. The expressed protein was 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 single-copy gene.
Capelli
N
N
Laboratory of General Microbiology, University of Geneva, Switzerland.
van Tuinen
D
D
Ortega Perez
R
R
Arrighi
J F
JF
Turian
G
G
D000595:Amino Acid Sequence; D001483:Base Sequence; D002147:Calmodulin / Q000235:genetics*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009492:Neurospora crassa / Q000235:genetics*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment
D002147:Calmodulin; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D004247:DNA