唐鱼卵黄蛋白原的诱导、纯化与鉴定

为探讨利用雄性唐鱼(Tanichthys albonubes)卵黄蛋白原(Vitellogenin,Vtg)作为生物标志物检测环境雌激素的可行性,采用浸浴法用50μg·L-117β-雌二醇(E2)对雄性唐鱼进行染毒,30d后将鱼体整体匀浆进行常规聚丙烯酰胺凝胶电泳(Native-PAGE)以分析Vtg的产生;并采用Mg2+-EDTA选择性沉淀和Q Sepharose阴离子交换层析对Vtg进行分离纯化.结果表明,E2诱导下,雄性唐鱼产生了雌性特异蛋白Vtg,并且可在体内积累;利用Mg2+-EDTA选择性沉淀和Q Sepharose阴离子交换层析的两步纯化方法可分离纯化唐鱼整体匀浆Vtg;经Native-PAGE鉴定,确定唐鱼Vtg的分子量为440kDa.以上结果提示,雄性唐鱼Vtg可以作为环境雌激素监测的有效生物学标记物。

Induction, Purification and Identification of Vitellogenin from Tanichthys albonubes

In order to test if vitellogenin(Vtg) could be a valid biomarker for environmental estrogens,adult male Tanichthys albonubes were exposed to 50μg·L-1 17β-estradiol(E2).After 30 days induction,whole body homogenate(WBH) of T.albonubes was carried out by native polyacrylamide gel electrophoresis(Native-PAGE),and purified by Mg2+-EDTA sedimentation and anion exchange chromatography.Results showed that Vtg could be induced from male T.albonubes treated with E2 and accumulated in the body.Mg2+-EDTA sedimentation and anion exchange chromatography could be used to separate and identify the Vtg in the WBH of T.albonubes.The molecular weight of Vtg of T.albonubes was approximately 440kDa.The results indicated that Vtg of the adult male T.albonubes could be a valid biomarker for environmental estrogens.