| 尼罗罗非鱼卵黄脂磷蛋白的分离纯化与性质鉴定 |
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| 引用本文： | 单瑞后,王松,王骏,王军,汝少国.尼罗罗非鱼卵黄脂磷蛋白的分离纯化与性质鉴定[J].中国水产科学,2015,22(4):638-644. |
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| 作者姓名： | 单瑞后 王松 王骏 王军 汝少国 |
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| 作者单位： | 1. 中国海洋大学 海洋生命学院, 山东 青岛 266003; 2. 中国水产科学研究院 黄海水产研究所, 山东 青岛 266071; 3. 山东出入境检验检疫局, 山东 青岛, 266002 |
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| 基金项目： | 国家质检总局课题计划项目(2013IK196). |
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| 摘 要： | 采用Sephacryl S-300过滤层析和DEAE-Sepharose Fast Flow离子交换层析相结合的方法从尼罗罗非鱼(Oreochromis niloticus)成熟卵子匀浆液中分离纯化出了一种高分子量的蛋白。该蛋白能被Schiff试剂、甲基绿和苏丹黑B着色,Western blot显示能被金鱼卵黄脂磷蛋白(lipovitellin,Lv)多克隆抗血清特异性识别,在非变性条件下分子量约为560 k D,在SDS变性条件下分子量约为112 k D,结果表明分离纯化的蛋白是一种含有糖、磷、脂基团的蛋白,符合鱼类Lv的性质,且与金鱼Lv有免疫交叉反应,从蛋白的性质和免疫原性以及分子量大小等角度判断,本研究获得的高纯度蛋白为尼罗罗非鱼卵黄脂磷蛋白;纯化的罗非鱼Lv在反复冻融、37℃及60℃处理条件下均未出现降解,表明罗非鱼Lv比鱼类卵黄原蛋白(Vitellogenin,Vtg)更为稳定。研究结果为罗非鱼Lv抗体的制备奠定了基础。
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| 关 键 词： | 尼罗罗非鱼 卵黄脂磷蛋白 纯化 性质鉴定 |
| 修稿时间： | 2015/7/28 0:00:00 |
Purification, identification, and characterization of lipovitellin from tilapia (Oreochromis niloticus) |
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| SHAN Ruihou,WANG Song,WANG Jun,WANG Jun,RU Shaoguo.Purification, identification, and characterization of lipovitellin from tilapia (Oreochromis niloticus)[J].Journal of Fishery Sciences of China,2015,22(4):638-644. |
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| Authors: | SHAN Ruihou WANG Song WANG Jun WANG Jun RU Shaoguo |
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| Institution: | 1. Marine Life Science College, Ocean University of China, Qingdao 266003, China; 2.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; 3. Shandong Entry-Exit Inspection and Quarantine Bureau, Qingdao 26600 |
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| Abstract: | Lipovitellin (Lv)
is the major proteolytic product of vitellogenin (Vtg) in the ovary and has
been widely usedin studies of endocrine disruption. In this paper, a high
molecular weight protein was purified from the ovary extracts ofNile tilapia () using gel
filtration followed by ion-exchange chromatography. Results of nativepolyacrylamide
gel electrophoresis (PAGE), lipid staining with Sudan black B,
carbohydratestaining with Schiff reagent,and phosphorus staining with methyl
green identified the purified protein as a phosphoglycoprotein. Using westernblotting,
the protein was cross-reacted with the anti-goldfish Lv antisera. Native PAGE
determined the molecularweight of the protein to be approximately 560 kD, and a
single monomer of ~112 kD was detected by the sodium dodecylsulfate (SDS)-PAGE.
Based on this characterization, immunogenicity, and molecular weight, this
protein wasidentified as the Lv of the Nile tilapia. No degradation was
observed under conditions of multigelation or incubation at37 |
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| Keywords: | Oreochromis niloticus lipovitellin purification characterization |
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