Expression and characteristic of the Cu/Zn superoxide dismutase gene from the insect parasitizing fungus Cordyceps militaris.
A Cu/Zn-superoxide dismutase (SOD) gene was characterized from Cordycepes militaris by gene cloning, heterogeneous expression and function analysis. This 154-aa SOD (CmSOD) was deduced from a 465-bp gene cloned, showing 72-95 % sequence identity to Cu/Zn-SODs from other fungi. The deduced amino acid sequence of the cDNA is highly similar to Beauveria bassiana (95 %), Isaria tenuipes (94 %) and Claviceps purpurea (88 %), respectively. The SOD gene of C. militaris spin 589 bp and consisted of two introns and three exons. The CmSOD coding region sequence was inserted into plasmid pQE-30 in order to construct prokaryotic expression vector, then transformed into Escherichia coli M15 cells for expression, and a mass of rCmSOD was obtained by IPTG induction. The enzyme activity of the purified rCmSOD was approximately 714.48 U/mg after the assay. The study provided a way for in-depth research on the expression and regulation of the CmSOD, and the molecular mechanism of anti-oxidative effect in C. militaris.
Plant Biotechnology Research Center, School of Agriculture and Biology, Shanghai Jiao Tong University, No. 800, Dongchuan Road, Shanghai 200240, People's Republic of China. xuanweizhou@sjtu.edu.cn
Xuan-Wei
XW
Zhou
Xue-Fei
XF
Wang
Qi-Zhang
QZ
Li
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D032661:Cordyceps / Q000201:enzymology*; D004926:Escherichia coli; D005091:Exons; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D015870:Gene Expression; D007313:Insecta / Q000382:microbiology; D007438:Introns; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D040681:Structural Homology, Protein; D013482:Superoxide Dismutase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D005656:Fungal Proteins; D011994:Recombinant Proteins; D013482:Superoxide Dismutase