| Vitellogenin (Vtg) is usually produced in sexually active female fish. However, Vtg can be detected in male and juvenile fish exposed to estrogen or estrogen mimic chemicals. Therefore, Vtg is generally considered to be a good biomarker for environmental estrogens. Immunoassays are the most commonly used approaches for Vtg determination. These antibody-based methods have considerable advantages of sensitivity and selectivity, however, at the same time suffer from a long process incubation period and troublesome wash steps. Therefore, development of a novel, rapid, easy and accurate method for Vtg determination is necessary and important. Studies so far about Vtg as a biomarker for environmental estrogens were mostly limited to freshwater fish. Due to the increasing pollution in marine environment, Vtg of marine fish receives great concern. In this study, Pagrosomus major was chosen as an experimental animal. Vtg standard had been purified from plasma of Pagrosomus major. An analytical method for rapid determination of Vtg in fish plasma had been developed. Based on this, a study about Vtg in plasma of juvenile Pagrosomus major as a biomarker for environmental estrogens had been carried out. The detailed information is as follows.1. Induction, purification and initial characterization of Vtg from Pagrosomus majorVtg was largely induced in Pagrosomus major by one intraperitoneal injection of 17β-estradiol (E2). With anion exchange liquid chromatography, Vtg was effectively purified by a column filled with DEAE Sepharose Fast Flow resin. It was found that Vtg was present in plasma of E2 treated Pagrosomus major and control female fish, and absent from plasma of control male fish, which suggested that the protein was a female specific protein and could be induced by E2. The results of Native-PAGE showed that the Vtg of Pagrosomus major had two forms, with molecular weights around 570 kDa and 360 kDa, respectively. After being separated by SDS-PAGE, the...
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