藏绵羊肌红蛋白（Mb）基因克隆及肌肉Mb含量的测定

为探索藏绵羊适应青藏高原缺氧环境的分子机制,克隆了藏绵羊的肌红蛋白（Mb）基因,纯化了心肌Mb,同时比较了心肌和骨骼肌的Mb含量。用RT-PCR克隆了心肌Mb的cDNA,长度为640 bp,编码153个氨基酸。试验采用盐析、CM-Sephadex离子交换层析、Sephadex G-50凝胶层析等方法分离纯化了藏绵羊Mb,SDS-PAGE分析其分子量约为17kD。组织含量测定表明,藏绵羊心肌中Mb含量极显著高于骨骼肌。

The Tibetan Sheep Myoglobin:cDNA Cloning and Content in Muscles

To explore the molecular mechanisms for Tibetan sheep to adapt to the Qinghai-Tibetan Plateau hypoxic environment,the experiments were carried out to clone Tibetan sheep myoglobin(Mb) gene,to purify Mb from cardiac tissue,and to measure Mb content in cardiac muscles and skeletal muscles.Total RNA was isolated from Tibetan sheep cardiac muscle,RT-PCR was employed to clone the Mb cDNA of Tibetan sheep,resulting in a 640bp sequence,which codes for a peptide of 153 amino acids.Mb was purified from Tibetan sheep cardiac tissue by salting-out,CM-Sephadex exchange chromatography,and Sephadex G-50 gel chromatography.SDS-PAGE analysis showed that the molecular weight of the purified Mb was approximately 17 kD.Mb contents in cardiac tissues were significantly higher than skeletal muscles in Tibetan sheep.