摘要
用山羊IgG注射欧洲鳗鲡背部肌肉后,采用山羊IgG-琼脂糖亲和层析柱对欧鳗血清免疫球蛋白(IgM)进行分离纯化,并通过SDS-PAGE电泳对纯化后的IgM进行检测,同时制备兔抗欧鳗IgM血清,运用间接ELISA检测正常欧鳗血清(第0天)以及山羊IgG注射欧鳗后第7、14、21、28、35、42天的血清抗体水平,同时利用荧光定量PCR测定了第0天至第28天的欧鳗肾脏、脾脏和鳃中IgM基因表达水平变化.结果表明:纯化后的欧鳗IgM经SDS-PAGE检测含有一条分子质量约68 ku的重链,以及分子质量分别为26 ku和28 ku的两条轻链,获得的兔抗欧鳗IgM血清效价达1/51 200;欧鳗血清抗体水平在第28天达到峰值,脾脏IgM基因表达水平在第14天达到峰值,肾脏和鳃均在第21天达到峰值,血清抗体水平以及三种组织器官IgM基因表达水平的变化规律均为先升高、达到峰值后再降低.
Adult European eels, Anguilla anguilla, were immunized by intramuscular injection with goat IgG and then the purified IgM was isolated from the serum using goat IgG-agarose affinity chromatography. The purified eel IgM was used to immunize rabbits to produce anti-eel IgM antibody. Results of SDS-PAGE electrophoresis showed that the heavy chain of the serum IgM was 68 ku in size and there were two light chains with the molecular weight of 26 ku and 28 ku respectively. The titers of anti-eel IgM antiserum were 1：51 200. The antibody levels of the normal serum （day 0） and the immuned serum at day 7, 14, 21, 28, 35, 42 post injection was assayed by ELISA. The abundance of IgM transcripts in the kidney, spleen and gills in European eels from day 0 to day 28 were measured by real-time RT-PCR. The highest antibody ELISA value in eels was reached at day 28 post injection. The highest level of IgM gene in the spleen, kidney and gills appeared respectively at day 14, day 21 and day 21 post injection. The model of the time course changes of the serum antibody levels and the abundance of IgM transcripts in the kidney, spleen and gills in European eels were the same, increasing to the peak and then decreasing gradually.
出处
《集美大学学报（自然科学版）》
CAS
2009年第4期333-338,共6页
Journal of Jimei University：Natural Science
基金
农业部公益性行业(鳗鱼)科研专项项目(nyhyzx07-043)
福建省自然科学基金资助项目(B0810032)
福建省高校水产科学与食品安全重点开放实验室基金(2008J203)
集美大学博士启动基金(ZQ2008013)
