人锰超氧化物歧化酶cDNA的克隆、测序及表达

Cloning, Sequencing and Expression of Human Manganese Superoxide Dismutase Gene cDNA.

用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）以人肝细胞总ＲＮＡ为模板，扩增了人锰超氧化物歧化酶（ｈＭｎＳＯＤ）的ｃＤＮＡ片段，将此ｃＤＮＡ克隆到载体ｐＧＥＭ－Ｔ中对重组质粒进行限制酶切分析和序列测定，确定为含ｈＭｎＳＯＤｃＤＮＡ的重组质粒将该ｈＭｎＳＯＤｃＤＮＡ重组到表达载体ｐＢＶ２２０内，重组质粒在大肠杆菌ＤＨ５－α中表达ｈＭｎＳＯＤ，表达产物占菌体总蛋白的１４％。

The cDNA encoding human manganese superoxide dismutase was amplified from the human liver total RNA by RT-PCR, ligated into pBV220. The cloned gene was analyzed by restriction enzymes EcoR l, Sal Ⅰ, and BamH Ⅰ. The sequence of the cloned gene was determined. hMnSOD was induced and expressed upon temperature shift from 32℃ to 42℃.Mn2+ supplementation in the bacterial growth media resulted in about 3-fold increasing of SOD activity. The special protein expressed accounts for 14% of the total protein of the bacteria.