Molecular cloning of a cDNA encoding calmodulin from Neurospora crassa

FEBS Letters

Perez, Ruben Ortega¹  Arrighi, Jean-F.¹  Capelli, Nicolas¹  Turian, Gilbert¹  van Tuinen, Diederik¹ 

[1] Laboratory of General Microbiology, University of Geneva, Sciences III, CH-1211 Geneva 4, Switzerland

关键词: Neurospora crassa;    Calmodulin;    Nucleotide sequence;    cDNA expression;    CaM;    calmodulin;    SDS-PAGE;    sodium dodecylsulfate polyacrylamide gel electrophoresis;    IPTG;    isopropyl β-d-thio-galacto-pyranoside;    PMSF;    phenylmethylsulfonyl fluoride;    EGTA;    ethylene glycol-bis(β-aminoethyl ether) N;    N;    N';    N'-tetraacetic acid;    EDTA;    ethylenediaminetetraacetic acid;   

DOI  :  10.1016/0014-5793(93)80622-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A full-length cDNA encoding Neurospora crassa calmodulin was isolated from λZAP 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.

【 授权许可】

Unknown