Genetic variation of disease resistance in fish

本研究对克隆的牙鲆生物防御和免疫相关基因的cDNA进行了克隆、测序和部分特征分析。这些cDNA是转铁蛋白，补体成分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δ，IgM，IgD，IG轻链，IG κ链，抗病毒蛋白Mx，趋化因子受体，干扰素诱导蛋白56 K，溶菌酶，本研究建立了一种将外源基因导入鱼类的有效方法，并在转基因鱼类系统中开发了活性启动子。水合精子电穿孔效果显著，获得的转基因率是普通电穿孔法的3倍。另外，我们还发现青鳉B-actin启动子在大部分组织中都有活性，而鳟鱼vasa启动子仅在转基因虹鳟的生殖细胞中有活性。使用它们的单克隆抗体，可以用磁性细胞分离系统进行细胞分离。分离的中性粒细胞显示非特异性细胞毒性以及吞噬作用。并且这种细胞毒活性与环境温度相适应。证明了鲤鱼血小板的聚集能力。这些结果为鱼类免疫系统的研究提供了新的信息。我们对英国养殖业的主要寄生虫病--鲑Leopeophtheirus salmonis进行了RAPD和核糖体RNA基因分析，结果表明，来自苏格兰沿海不同地区的鲑Leopeophtheirus salmonis具有不同的种群。

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.