剑尾鱼卵黄脂磷蛋白的纯化及免疫分析  

Purification and immune analysis of lipovitellin from Xiphophorus helleri

作　　者：温茹淑[1] 方展强[1] 江世贵[2] 马广智[1] 徐杰[1] 姚静[1] 

机构地区：[1]华南师范大学生命科学学院 [2]农业部渔业生态环境重点开放实验室广东省渔业生态环境重点实验室中国水产科学研究院南海水产研究所,广东广州510300

出　　处：《水产学报》2007年第5期647-654,共8页Journal of Fisheries of China

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(2004B40101015)

摘　　要：　采用Sephacryl S-300凝胶过滤层析柱和HiTrap Q阴离子交换柱从卵黄生成期的雌性剑尾鱼（Xiphophorus helleri）卵巢组织提纯了卵黄脂磷蛋白（Lv）。已确定被纯化的剑尾鱼Lv在Native-PAGE（4％～7．5％）电泳中分子量为530kD左右。Native-PAGE（4％～7．5％）电泳后的凝胶分别进行糖蛋白染色、脂蛋白染色及磷蛋白染色。结果表明，剑尾鱼Lv是一种富含糖、脂、磷的蛋白。用纯化的剑尾鱼Lv免疫大白鼠获得鼠源多克隆抗血清。用免疫双扩散的方法测定Lv抗血清的效价为1：32。Western-blotting检测显示抗血清的特异性较好；卵黄蛋白原（Vtg）和Lv抗血清之间存在明显的免疫交叉反应，表明Vtg和Lv两者具有相同的免疫原性。免疫双扩散检测表明，抗Lv血清表现出明显的雌性特异性和种的特异性。Lipovitellin （Lv） was purified from ovaries of mature female swordtail fish （ Xiphophorus helleri） by gel filtration with Sephacryl S-300 HR and anion exchange chromatography with HiTrap Q. The results indicated that the purified Lv appeared approximately 530 kD in native polyacrylamide gel electrophoresis （PAGE）. Lv was characterized as a phospholipoglycoprotein by Native-PAGE and staining of gels for carbohydrates, lipids and phosphorus. Polyclonal antibodies against the purified Lv from swordtail fish were produced in the mice. Double immunodiffusion showed that the determination of the titre for anti-Lv sera was 1 ： 32. Western-blotting analyses demonstrated that anti-Lv sera have specificity and Lv has common antigenicity with vitellogenin（Vtg） while purified Vtg and Lv had cross-reacted obviously with the anti-Lv sera. Double immunodiffusion showed that anti-Lv sera have sex-specificity and species-specificity.

关 键 词：剑尾鱼 卵黄脂磷蛋白 纯化 免疫分析 

分 类 号：Q579.1[生物学—生物化学] S917[农业科学—水产科学]