中文摘要：

腹腔注射17β-雌二醇（E2），使瓦氏黄颡鱼雄鱼在7天内产生卵黄蛋白原（Vtg）。采用凝胶过滤和离子交换两种层析技术，从E2诱导的雄性瓦氏黄颡鱼血浆中分离、纯化出Vtg，采用糖、磷、脂蛋白染色技术证明分离、纯化的蛋白为Vtg，该Vtg在非变性条件下分子量约为240kDa，在SDS变性条件下分子量约为143kDa。纯化的瓦氏黄颡鱼Vtg经检测显示可能含有类胡萝卜素，但没有二硫键，对热相对稳定。利用纯化的瓦氏黄颡鱼Vtg，制备了兔抗瓦氏黄颡鱼Vtg多克隆抗血清。用双向免疫扩散法测得抗血清的纯度较高，效价为1：32；Western blotting检测显示抗血清的特异性较好。以瓦氏黄颡鱼Vtg多克隆抗血清为抗体，以纯化的瓦氏黄颡鱼Vtg为抗原，建立了间接酶联免疫吸附反应（ELISA）方法检测瓦氏黄颡鱼体内Vtg的含量，标准曲线线性部分的线性方程为Y=0．099x＋0．4529（R^2=0．9327），该方法检测的灵敏度为15．6ng／ml，工作范围为31．2-4000ng／ml，在此范围内，标准曲线具有良好的线性。

英文摘要：

7-day after intraperitoneal injection of 17 β -estradiol （E2）, male Pelteobagrus vachelli produced vitellogenin （Vtg）; and later the Vtg from the E2 treated P. vachelli plasma was isolated and purified by gel filtration and ion-exchange chromatography. With phosphor-, lipo- and glycol-protein staining methods, we verified this protein as Vtg, in molecular weight of about 240kDa detected by Native-PAGE. In SDS-PAGE, the Vtg broke into 2 same subunits, each at 143kDa. The purified Vtg contained carotenoid of non-disulfide bond, relatively stable to heat. To make use of purified Vtg, we prepared polyclonal antiserum against P. vachelli Vtg. Double immunodiffusion determined that the titre for Vtg antisera was 1 ： 32; and western-blotting demonstrated that polyclonal antiserum had preferably specific effect. An indirect com- petitive enzyme-linked immunosorbent assay （ELISA） has been then established for detecting the Vtg. The technique was developed using Vtg-resistant antiserum as antibody and Vtg as antigen in working range of 31.2--4000ng/ml, and the sensitivity at 15.6ng/ml. The equation linear part of a typical ELISA calibration curve is y =0.099x ＋ 0.4529 （R^2 = 0.9327）, which shows good linearity in the working range.