泰山螭霖鱼卵黄脂磷蛋白的纯化及免疫原性分析  

Purification and Immunoassay Study of Lipovitellin from Varicorhinus Macrolepis

作　　者：李盼 李平江 林山 李永琪 林贞贤[2] LI Pan;LI Ping-jiang;LIN Shan;LI Yong-qi;LIN Zhen-xian(School of Life Sciences,Shandong Agricultural University,Taian,Shandong 271000;School of Biology and Brewing Engineering,Taishan University,Taian,Shandong 271021)

机构地区：[1]山东农业大学生命科学学院,山东泰安271000 [2]泰山学院生物与酿酒工程学院,山东泰安271021

出　　处：《泰山学院学报》2024年第6期47-53,共7页Journal of Taishan University

基　　金：泰安市科技创新发展项目“泰山螭霖鱼气调保活运输技术研究”(2022NS112)。

摘　　要：采用阴离子交换层析结合超滤方法从性成熟的雌性泰山螭霖鱼(Varicorhinus macrolepis)卵匀浆液中纯化出卵黄脂磷蛋白(Lipovitellin,Lv),该蛋白的天然分子量约为525 kD,在SDS-PAGE电泳显示为分子量为112.5、103.6和92.4 kD三个亚基。利用纯化的Lv作为抗原制备了兔抗螭霖鱼Lv多克隆抗血清。双向免疫扩散实验和Western blot结果显示,兔抗螭霖鱼Lv血清与螭霖鱼卵黄蛋白原(Vitellogenin,Vtg)、斑马鱼(Danio rerio)Lv和唐鱼(Tanichthys albonubes)Lv均能发生很好的免疫交叉反应。本研究结果为螭霖鱼Vtg准确定量检测方法的开发奠定了基础。In this study,ion-exchange chromatography and ultrafiltration were used to purify lipovitellin(Lv)from ovarian homogenates of Varicorhinus macrolepis.The natural molecular weight of the protein was about 525 kD,and three subunits with molecular weights of 112.5,103.6 and 92.4 kD were detected by SDS-PAGE.The rabbit polyclonal antiserum against Lv of Varicorhinus macrolepis was prepared.The results of the double immunodiffusion agar test and western blot showed that antiserum had good cross reaction with vitellogenin of Varicorhinus macrolepis,zebrafish(Danio rerio)Lv,and Tanichthys albonubes Lv.Results of the present study had laid foundation for the development of an accurate quantitative detection method for vitellogenin in Varicorhinus macrolepis.

关 键 词：泰山螭霖鱼 卵黄脂磷蛋白 纯化 免疫分析 

分 类 号：S917[农业科学—水产科学]