摘要
目的 研究17p雌二醇（E2）暴露对雄性剑尾鱼（Xiphophorus helleri）卵黄蛋白原（vitellogenin，Vtg）诱导作用作为环境风险评价（ERA）的有效生物学标记的可行性。方法 以E2诱导的剑尾鱼雄性个体的整体匀浆液为材料，采用Sephaeryl S-300凝胶过滤层析柱和Q Sepharose阴离子交换柱从剑尾鱼体内提纯Vtg。结果确定了被纯化的剑尾鱼Vtg在4%-7．5%Native PAGE电泳中相对分子质量为540×l0^3 . 4%-7．5%Native PAGE电泳后的凝胶分别利用苏丹黑B进行脂蛋白染色、高碘酸．Schiff试剂进行糖蛋白染色和甲基绿进行磷蛋白染色，表明剑尾鱼Vtg是一种富含糖、脂、磷的蛋白。结论表明雄性剑尾鱼卵黄蛋白原的诱导变化可作为环境风险评价（ERA）的有效生物学标记。
Objective To explore if induction of vitellogenin （Vtg） by 17β-estradiol in male swordtail fish （Xiphophorus hel/er/） can be used as a valid biomarker for environmental risk assessment （ERA）. Methods Vtg was purified from whole body homogenate （WBH） of estradiol-17β treated （10μg/mL ambient freshwater for 21 days） male swordtail fish by gel filtration （Sephacryl S-300） and anion exchange chromatography （Q Sepharoso）, respectively. Results Vtg appeared to exist as a homotrimer of approximately 540 ×10^3 in native polyacrylamide gel electrophoresis （PAGE）. Vtg was characterized as a phospholipoglycoprotein by native PAGE and staining of gels for phosphorus with methyl green,for lipids with Sudan black B, and for carbohydrates with periodic acid-Schifl reagent. It also indicated that the purified proteins resembled the Vtg in other teleost species, and therefore reasonably identified as swordtail fish Vtg. Conclusion The results indicated that Vtg in male swordtail fish could be considered as a valid biomarker for environmental risk assessment （ERA）.
出处
《中国实验动物学报》
CAS
CSCD
2007年第4期280-284,I0003,共6页
Acta Laboratorium Animalis Scientia Sinica
基金
农业部渔业生态环境重点开放实验室开放基金
广东省渔业生态环境重点实验室开放基金项目(2005-9)
广东省科技计划项目(编号:2004B40101015)资助
关键词
剑尾鱼
卵黄蛋白原
提纯
环境风险评价
Swordtail fish （ Xiphophorus helleri）
Vitellogenin
Purification
Environmental risk assessment
