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用于测定加利福尼亚大比目鱼(加利福尼亚牙鲆)卵黄蛋白原的酶联免疫吸附测定法的验证

Validation of an enzyme-linked immunosorbent assay for measuring vitellogenin in California halibut (Paralichthys californicus).

作者信息

Vázquez-Boucard Celia, Burgos-Aceves Mario, Arcos-Ortega Fabiola, Anguiano-Vega Gerardo

机构信息

Aquatic Organisms Reproduction and Toxicogenomic Laboratory, Department of Environmental Management and Conservation, Centro de Investigaciones Biológicas del Noroeste, La Paz, Baja California Sur, México.

出版信息

Environ Toxicol Chem. 2008 Jul;27(7):1614-20. doi: 10.1897/06-368.

Abstract

Vitellogenin (VTG) is the major protein present in the plasma of females undergoing oogenesis. In males, the VTG gene normally is suppressed; however, synthesis of VTG can be induced by exposure to xenoestrogenic compounds. In the present study, an enzyme-linked immunosorbent assay was developed and validated to evaluate VTG levels in the California halibut (Paralichthys californicus). Vitellogenin and lipovitellin (LV) were identified in the plasma of 17 beta-estradiol-induced females and in the ovaries of wild females, to our knowledge for the first time. Purified VTG from the plasma of induced females was obtained, and polyclonal antibodies against the LV of mature female ovaries was prepared and their specificity assessed by Western blot analysis. At Bahía Magdalena, Baja California Sur, México, quantitative measurements of VTG in the plasma of female specimens were made during one reproductive cycle.

摘要

卵黄蛋白原(VTG)是正在进行卵子发生的雌性动物血浆中的主要蛋白质。在雄性动物中,VTG基因通常受到抑制;然而,接触外源性雌激素化合物可诱导VTG的合成。在本研究中,开发并验证了一种酶联免疫吸附测定法,以评估加利福尼亚大比目鱼(Paralichthys californicus)的VTG水平。据我们所知,首次在17β-雌二醇诱导的雌性动物血浆和野生雌性动物卵巢中鉴定出卵黄蛋白原和脂卵黄蛋白(LV)。从诱导雌性动物血浆中获得了纯化的VTG,并制备了针对成熟雌性动物卵巢LV的多克隆抗体,并通过蛋白质印迹分析评估了它们的特异性。在墨西哥南下加利福尼亚州的马格达莱纳湾,在一个生殖周期内对雌性样本血浆中的VTG进行了定量测量。

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