Capelli N, van Tuinen D, Ortega Perez R, Arrighi J F, Turian G
Laboratory of General Microbiology, University of Geneva, Switzerland.
FEBS Lett. 1993 Apr 19;321(1):63-8. doi: 10.1016/0014-5793(93)80622-2.
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.
从λZAP II cDNA表达文库中分离出了编码粗糙脉孢菌钙调蛋白的全长cDNA。该开放阅读框编码一个由148个氨基酸残基组成的蛋白质，计算分子量为16,865道尔顿。利用定点诱变技术，将完整的cDNA连接到受trc启动子调控的细菌表达载体中，以便在大肠杆菌中表达粗糙脉孢菌钙调蛋白。根据其一些生化特性，发现表达的蛋白质与天然蛋白质相同。最后，对基因组DNA酶切片段的Southern分析表明，钙调蛋白由单拷贝基因编码。
