长鳍吻(鳍)免疫因子基因cDNA部分序列克隆及其组织表达差异

根据鲤科鱼类同源序列设计并合成了长鳍吻(鳍)(Rhinogobio ventralis)应激因子HSP70、抗体IgM、抑炎性因子IL-10和促炎性因子IL-1b基因特异引物,以长鳍吻(鳍)皮肤组织总RNA为模板,RT-PCR 扩增获得长度分别为412、463、520和217 bp的上述4种免疫因子基因cDNA部分序列.同时通过RT-PCR比较患小瓜虫病长鳍吻1715和对照组长鳍吻(鳍)皮肤和肠道组织中各免疫因子的表达差异,结果显示:在皮肤组织中,IgM和IL-10在感染组鱼体中表达量显著高于对照鱼(P<0.05), HSP 70和 IL-1b的表达则没有差异;在肠道组织中,IgM、IL-10和 IL-1b在感染组鱼体中表达量显著增高,HSP 70的表达量没有差异.本研究首次对长鳍吻(鳍)的免疫因子进行了分析,指出免疫因子IgM和IL-10在鱼体中免疫应答反应中较为灵敏,为养殖过程中长鳍吻(鳍)应激监测方面奠定了理论基础.

Cloning and tissue expression of some immune factor genes in Rhinogobio ventralis

In order to get the partial cDNAsequences of some immune factors gene in Rhinogobio ventralis, including stress factor HSP70, IgM antibody, anti-inflammatory factor IL-10 and proinflammatory factor IL-1b gene specific primers of R.ventralis were designed and synthesized according to the homologous sequence of Cyprinidae.Four different immune factor gene cDNA sequences were obtained by RT-PCR amplification, which respectively for length 412, 463, 520 and 217 bp.The quantitative (RT-PCR)method was used to detect the mRNA differential expression of HSP 70, IgM, IL-10 and IL-1b in the skin and gut tissues of R.ventralis infected by ichthyophthiriasis group and control group.The results showed that the expressions of IgM and IL-10 in skin tissues of the infected group were significantly higher than that of control group.The expression of the IgM、IL-10 and IL-1b in the gut tissues of infected group were significantly higher than that of control group.However, no significant difference was shown in the expression of HSP 70 between two groups.This is the first report to analyze the immune factors of R.ventralis, and the highlight shows that immune factors of IgM and IL-10 in fish immune response are more sensitive.All results will lay the theoretical foundation for monitoring the stress response of R.ventralis in recirculating aquaculture system.