中国水产科学Issue(4)：638-644,7.DOI:10.3724/SP.J.1118.2015.140442
尼罗罗非鱼卵黄脂磷蛋白的分离纯化与性质鉴定
Purification, identification, and characterization of lipovitellin from tilapia (Oreochromis niloticus)
摘要
Abstract
Lipovitellin (Lv) is the major proteolytic product of vitellogenin (Vtg) in the ovary and has been widely used in studies of endocrine disruption. In this paper, a high molecular weight protein was purified from the ovary extracts of Nile tilapia (Oreochromis niloticus) using gel filtration followed by ion-exchange chromatography. Results of native polyacrylamide 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 western blotting, the protein was cross-reacted with the anti-goldfish Lv antisera. Native PAGE determined the molecular weight of the protein to be approximately 560 kD, and a single monomer of ~112 kD was detected by the sodium do-decyl sulfate (SDS)-PAGE. Based on this characterization, immunogenicity, and molecular weight, this protein was identified as the Lv of the Nile tilapia. No degradation was observed under conditions of multigelation or incubation at 37℃ or 60℃; thus, the purified Nile tilapia Lv exhibited a relatively higher stability than vitellogenin. To our know-ledge, this is the first report of the purification and characterization of the Nile tilapia Lv. Results of the present study provide a theoretical foundation for the manufacture of polyclonal antisera against Nile tilapia Lv which could then be used to detect vitellogenin induction in tilapia.关键词
尼罗罗非鱼/卵黄脂磷蛋白/纯化/性质鉴定Key words
Oreochromis niloticus/lipovitellin/purification/characterization分类
农业科技引用本文复制引用
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,(4):638-644,7.基金项目
国家质检总局课题计划项目(2013IK196) （2013IK196）
