唐鱼卵黄脂磷蛋白的纯化鉴定与免疫原性分析  

Purification and Immune Analysis of Lipovitellin from Tanichthys alonues

作　　者：姚静[1] 方展强[1] 徐杰[1] 杨丽丽[1] 张灶桂[1] 唐家宁[1] 

机构地区：[1]华南师范大学生命科学学院

出　　处：《应用与环境生物学报》2008年第1期69-73,共5页Chinese Journal of Applied and Environmental Biology

基　　金：农业部渔业生态环境重点开放实验室开放基金项目(No2005-9);广东省科技计划项目(No2004B40101015)资助~~

摘　　要：采用Native-PAGE和SDS-PAGE方法从性成熟雌性唐鱼（Tanichthys albonubes）卵巢组织提纯了卵黄脂磷蛋白（Lv）．已确定被纯化的唐鱼Lv在Native-PAGE（4％～7．5％）电泳中分子量（Mr）为314×10^3．用纯化的唐鱼Lv免疫大白鼠获得鼠源多克隆抗血清．以Native-PAGE和SDS-PAGE制备的唐鱼Lv及裂解后的组分作为抗原所制备的抗血清，与经17β-雌二醇（E2）诱导的雄性唐鱼、去除卵巢的雌性唐鱼以及唐鱼卵巢的匀浆液能发生免疫反应，并且印迹位置与雌性特异蛋白卵黄蛋白原（Vtg）的位置相当．但是两种血清和未经E2诱导的雄鱼整体匀浆液并无反应，显示Lv及其大分子亚基的抗血清与Vtg和Lv两种蛋白足特异的．结果表明，唐鱼Lv裂解组分的抗血清可用于检测唐鱼的Vtg．Lipovitellin （Lv） was purified from the ovaries of mature female white mountain minnow （ Tanichthys albonubes） by native - polyacrylamide gel electrophoresis （ Native - PAGE） and sodium dodecylsulfate - polyacrylamide gel electrophoresis （SDS- PAGE）. The results indicated that the Lv appeared to exist as homotrimer of approximately 314×10^3 in Native - PAGE （4% -7.5% ）. Polyclonal antiserum against Lv was raised in rat. Native- PAGE followed by Western blotting was performed to analyze the specificity of the antiserum and immunological similarities between Lv and vitellogenin （Vtg）. Only one protein band in the whole body homogenate from 17β-estradiol （E2 ） treated male T. albonubes and homogenate from ovaties respectively was reacted with the antiserum. No protein band from untreated males was reacted with the anti-Lv serum and anti-high molecular weight （HMW） subunit of Lv serum. The results also indicated that anti - HMW subunit of Lv could be used for immune analysis and measuring Vtg. Fig 3, Ref 19

关 键 词：唐鱼 卵黄脂磷蛋白 纯化 免疫分析 

分 类 号：Q51[生物学—生物化学]