美国红鱼卵黄原蛋白的分离纯化及电泳性质鉴定

Purificaton and Electrophoretic Comparison of Sciaenops ocellatus Vitellogenin

腹腔注射0.02mg/gBW17β-雌二醇,诱导美国红鱼1wk后取尾静脉血,离心分离得到的血浆经Sephacryl S-300 high resolution分子筛分离、纯化卵黄原蛋白.常规聚丙烯酰胺凝胶电泳的考马斯亮蓝染色和糖、磷、脂的特征性基团染色结果表明,17β-雌二醇诱导美国红鱼产生了两种形式的卵黄原蛋白,以Native-PAGE方法计算出美国红鱼两种卵黄原蛋白的分子量分别为344×103和247×103,以SDS-PAGE方法得出卵黄原蛋白3种亚基的分子量为152×103、133×103和118×103.分子筛凝胶过滤能较好地分离纯化美国红鱼血浆中的卵黄原蛋白.

Vitellogenin （ VTG ） was induced in red drum by intraperitoneal injections of 17β-estradiol. After one week induction, blood was collected from caudal vessel and then centrifuged. Purification of the VTG was performed by gel permeation chromatography of Sephacryl S-300 high resolution. After plasma native-PAGE, the gel was stained by the methods of CBB G250-, phospho-, lipo- and glyco-staining, respectively. The result of native-PAGE showed after one week induction, red drum synthesized two kinds of vitellogenins, and the molecular weights of vitellogenins were 344 ×10^3 and 247 ×10^3. The purified vitellogenin protein was appeared as 152 ×10^3 , 133 ×10^3 and 118 ×10^3 in SDS - PAGE under reducing condition. The two kinds of vitellogenins in plasma were effectively purified by gel permeation chromatography of Sephacryl S-300 high resolution. Fig 8, Ref 17