鲟鱼GtH放射免疫测定方法的建立及催产前后史氏鲟血清GtH含量的变化
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

国家科技部“十五”863现代农业技术主题(2001AA243061,2004AA243060);北京市科学技术委员会(9548304000)


Setting up radioimmunoassay for sturgeon gonadotropin GtH and assay of Amur sturgeon serum GtH after inducing ovulation
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    用蛋白质快速液相层析(FPLC)分离俄罗斯鲟鱼脑垂体促性腺激素(GtH),共获得GtH纯化蛋白4.6 mg。用SDS-PAGE变性胶电泳图谱初步分析纯化蛋白,用反相高效液相色谱(rpHPLC)分离出两个亚基,质谱测定α亚基分子量:15 603 ku,两个β亚基分子量分别为14 338 ku和14 694ku。制备了鲟鱼GtH的兔抗血清多克隆抗体,采用氯胺T法用125I标记抗原,用羊抗兔γ球蛋白做二抗,建立鲟鱼GtH的放射免疫测定方法。测定了催产前后不同时间史氏鲟血清中GtH含量的变化,统计分析表明成功排卵的雌鱼比未排卵和阴性对照组在催产后12 h和16 h GTH分泌量升高显著,最高达到234 ng·mL-1;催产效应时间与催产后血清中GtH的含量紧密相关。

    Abstract:

    Gonadotropin (GtH) protein was purified from the pituitary of Russian sturgeon using fast protein liquid chromatograph (FPLC). A total of 4.6 mg of Russian sturgen GtH in dry weight was obtained. The purified protein was analyzed by SDS-PAGE electrophoresis. Two subunits of GtH were separated by reversed-phase high-performance liquid chromatography (rpHPLC). The molecular weight was determined through mass spectrograph. The α subunit was 15 603 ku and the two β-subunits were 14 338 ku and 14 694 ku respectively. Rabbit anti-sturgeon GtH polyclone antibody was prepared; purified GtH was labeled by 125I using chloramines T as labeled antigen; then the sturgeon GtH radioimmunoassay (RIA) method was set up using the goat anti-rabbit γ-globulin as the second antibody . After inducing ovulation, changes of serum GtH levels were detected. The secretion peak of female fish ovulated successfully was sustained from 12 to 16 hours after first inducing injection and the serum GtH levels were increased significantly compared with control group and un-ovulated fish. The effect time of LHRH-A was related close to the serum GtH levels.

    参考文献
    相似文献
    引证文献
引用本文

胡红霞,刘晓春,朱华,张勇.鲟鱼GtH放射免疫测定方法的建立及催产前后史氏鲟血清GtH含量的变化[J].水产学报,2008,32(6):817~824

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2007-11-28
  • 最后修改日期:2008-05-28
  • 录用日期:2008-09-17
  • 在线发布日期: 2008-11-13
  • 出版日期: