曹琦琦, 王玉红, 仲小芳, 陈常仪, 王安利. 尼罗罗非鱼IgT重链的蛋白表达及表达分析[J]. 华南师范大学学报(自然科学版), 2018, 50(4): 62-67. doi: 10.6054/j.jscnun.2018080
引用本文: 曹琦琦, 王玉红, 仲小芳, 陈常仪, 王安利. 尼罗罗非鱼IgT重链的蛋白表达及表达分析[J]. 华南师范大学学报(自然科学版), 2018, 50(4): 62-67. doi: 10.6054/j.jscnun.2018080
CAO Qiqi, WANG Yuhong, ZHONG Xiaofang, CHEN Changyi, WANG Anli. The Protein Expression and Expression Pattern Analysis of IgT Heavy-Chain in Nile Tilapia (Oreochromis niloticus)[J]. Journal of South China Normal University (Natural Science Edition), 2018, 50(4): 62-67. doi: 10.6054/j.jscnun.2018080
Citation: CAO Qiqi, WANG Yuhong, ZHONG Xiaofang, CHEN Changyi, WANG Anli. The Protein Expression and Expression Pattern Analysis of IgT Heavy-Chain in Nile Tilapia (Oreochromis niloticus)[J]. Journal of South China Normal University (Natural Science Edition), 2018, 50(4): 62-67. doi: 10.6054/j.jscnun.2018080

尼罗罗非鱼IgT重链的蛋白表达及表达分析

The Protein Expression and Expression Pattern Analysis of IgT Heavy-Chain in Nile Tilapia (Oreochromis niloticus)

  • 摘要: IgT(Immunoglobulin T)作为新型免疫球蛋白在粘膜免疫中具有重要作用. 文中根据尼罗罗非鱼IgT基因序列构建原核表达载体获得IgT重组蛋白,相对分子量约75 kDa,并制备出效价较高的小鼠抗罗非鱼IgT的多克隆抗体. 荧光定量PCR检测证明:健康尼罗罗非鱼中IgT在多种组织中都有表达,其中脾脏、皮肤、肠和鳃中表达量相对较高. 无乳链球菌感染后,皮肤和头肾中IgT mRNA的相对表达量分别在12 h和4 d显著上调. 免疫组化结果显示:IgT蛋白在头肾和皮肤中均有少量分布,在无乳链球菌刺激后,在皮肤中出现大量分布. 以上结果表明:IgT在尼罗罗非鱼皮肤粘膜免疫中具有重要作用.

     

    Abstract: As a new type of immunoglobulin, IgT (Immunoglobulin T) plays an important role in mucosal immunity. The prokaryotic expression vector of Nile tilapia IgT was constructed according to its gene sequence to obtain the IgT recombinant protein, weighing 75103 Da approximately, and the mouse polyclonal antibody against IgT with high titer was prepared. Expression analysis showed that IgT was expressed in various tissues, and the relatively high expression was in spleen, skin, intestine and gills. The relative expression of IgT mRNA in the head kidney and skin was significantly up-regulated in 12 h and 4 d after the infection of S.agalactiae. The results of immunohistochemistry revealed that IgT protein had a small amount of distribution in the head kidney and skin; however, there was an obvious up-regulation in the skin after the stimulation of S.agalactiae. Taken together, the results of this study indicate that IgT plays a main role in mucosal immunity of Nile tilapia.

     

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