Genetic variation of disease resistance in fish

我们克隆、测序并部分鉴定了几个克隆的牙鲆的生物防御和免疫相关基因。这些基因包括转铁蛋白、补体C3、C7、C8和C9、补体调节因子、干扰素调节因子、T细胞受体α和Delta、1gm、IGD、Ig轻链、Ig kappa链、抗病毒蛋白Mx、趋化因子受体、干扰素诱导蛋白56K、溶菌酶等。水合精子电穿孔非常有效，转化率是常规电穿孔方法的3倍以上。此外，我们还发现Medaka B-肌动蛋白启动子在大多数组织中都非常活跃；然而，鲑鱼Vasa启动子只在转基因虹鱼的生殖细胞中活性。我们制备了抗鲤鱼白细胞，特别是中性粒细胞和血小板的单抗。利用它们的单抗，可以用磁性细胞分离系统进行细胞分离。分离的中性粒细胞表现出非特异性的细胞毒性和吞噬作用。这种细胞毒活性适应了环境温度。证明了鲤鱼血小板的聚集能力。对英国水产养殖业的主要寄生虫病鲑鱼进行了随机扩增多态性DNA和核糖体RNA基因分析，显示了苏格兰沿海不同地区的不同种群。

We have cloned, sequenced and partially characterized several biodefenese and immune *elated gene cDNA from cloned Japanese flounder. These cDNA were transferrin, complement component C3, C7, C8, and C9, complement regulatory factor, interferon regulator factor, T cell receptor alpha and delta, 1gM, IgD, Ig light chain, Ig kappa chain, antivirus protein Mx, Chemokine receptor, interferon inducible protein 56K, lysozyme, and so on.We have developed an effective method to introduce foreign gene into fish and have exploited the active promoters in transgenic fish systems. The electroporation of hydrating sperm was very effective and the obtained transgenic rate was 3 times higher than the ordinal electroporation method. Also, we have showed that Medaka B-actin promoter was very active in most of the tissue ; however, trout vasa promoter was active only in germ cells in transgenic rainbow trout.We made monoclonal antibodies against carp leukocytes, especially neutrophilic granulocytes and thrombocytes. Using their monoclonal antibodies, cell separation could be carried out with magnetic cell separation systems. Neutrophilic granulocytes separated showed non-specific cytotoxity as well as phagocvtosis. And this cytotoxic activity adapted to environmental temperatures. Aggregation capacity of carp thrombocytes was proved. These results provided some new informations out fish immune system clearly.We carried out RAPD and ribosomal RNA gene analysIs on Leopeophtheirus salmonis, the major parasitic disease in UK aquaculture, which showed distinct populations from different areas around the Scotland coast.