利用弱阴离子交换色谱,从雌激素17β-雌二醇诱导的真鲷鱼血清中分离纯化出卵黄蛋白原粗组分,以超滤法进一步除去杂质蛋白,并脱盐浓缩,获得纯度较高的卵黄蛋白原(Vtg),两级纯化得到的Vtg经常规聚丙烯酰胺凝胶电泳(Native-PAGE)分析,证实真鲷鱼Vtg有三种形式,分子量分别为665kDa,635kDa和355kDa;变性聚丙烯酰胺凝胶电泳(SDS-PAGE)分析结果表明,它们由185kDa左右的蛋白亚基组成.With ultrafiltration and anion exchange liquid chromatography,a two-step method had been developed for purification of vitellogenin(Vtg) from the serum of Pagromus major induced by 17β-estradiol(E_(2)).The(results) showed that most impurities could be removed during the first step by weak anion exchange liquid(chromatography) and further clean-up through ultrafiltration.In Native-PAGE,the Vtg purified by the two steps showed three forms,with molecular weights approximately 665kDa,635kDa and 355kDa,respectively.The result of SDS-PAGE indicated that the molecular weight of Vtg monomers was about 185kDa.High resolution anion exchange high performance liquid chromatography was applied to analyze the Vtg purified from the two-step method,and approved the high purity.The purified Vtg could be used for further analysis.国家自然科学基金(20277030)资助项
