食品科学 ›› 2008, Vol. 29 ›› Issue (5): 78-81.

• 基础研究 • 上一篇    下一篇

铜诱导华溪蟹金属硫蛋白的分离纯化

马文丽, 王兰   

  1. 山西大学生命科学与技术学院; 山西大学生命科学与技术学院;
  • 出版日期:2008-05-15 发布日期:2011-08-26

Isolation and Purification of Metallothionein from Sinopotamon henanense Induced with Copper

 MA  Wen-Li, WANG  Lan   

  1. School of Life Science and Technology,Shanxi University
  • Online:2008-05-15 Published:2011-08-26

摘要: 采用体外铜暴露法对河南华溪蟹(Sinopotamon henanense)进行处理,诱导金属硫蛋白(metallothionein,MT)的高效表达。取肝胰腺组织匀浆、冷冻离心,采用丙酮沉淀粗提(50%~80%),然后将MT粗提液依次经过Sephadex G-50凝胶过滤层析、DEAE-Sepharose Fast Flow离子交换层析、透析脱盐、冷冻干燥,分离纯化MT,并进行洗脱峰的紫外光谱扫描分析、SDS-聚丙烯酰胺凝胶电泳分析(SDS-PAGE)及质谱分析。结果表明:得到了纯度较高的华溪蟹MT,主要以单体及二聚体形式存在,其分子量为6882D。

关键词: 铜, 河南华溪蟹, 金属硫蛋白, 分离纯化

Abstract: The freshwater crabs(Sinopotamon henanense) were exposed with 100 mg/L Cu(as CuSO4·5H2O) to induce synthesizing of metallothionein(MT).Then the crabs were dissected and the hepatopancreas were taken out,homogenized and centrifugalized.The supernatant obtained were purified primarily using acetone precipitation(50 %~80 %),followed by chromatography treatment on columns Sephadex G-50 and DEAE sepharose fast flow,then dialyzed to desalt.According to SDS-PAGE analysis,UV absorbance spectrometry analysis and mass spectrometry analysis,high purity MT is obtained,and its molecular weight is 6882 D.

Key words: copper, Sinopotamon henanense, metallothionein, isolation and purification