张锐,王子昂,汤俊宇,杨林蔚

• 1:广东海洋大学食品科技学院/现代生化中心
• 2:广东海洋大学深圳研究院
• 3:中山大学生命科学学院有害生物控制与资源利用国家重点实验室
• 4:南方海洋科学与工程广东省实验室(珠海)

摘要(Abstract)：

【目的】克隆凡纳滨对虾(Litopenaeus vannamei)一种新的含酪氨酸酶结构域血蓝蛋白(Tyrosinase-domain containing hemocyanin,TDCH)，研究其免疫功能。【方法】根据NCBI上公布的tdch基因序列设计克隆引物，通过RT-PCR方法获得tdch的编码区；采用生物信息学方法分析tdch的结构特征；使用荧光定量PCR技术分析tdch在凡纳滨对虾12个组织中的表达，以及在副溶血弧菌(Vibrio parahaemolyticus)、金黄色葡萄球菌(Staphhylococcu saureus)、黑曲霉菌(Aspergillus niger)、白斑综合征病毒、脂多糖及聚肌苷酸-聚胞苷酸[Poly(I:C)]刺激后在鳃和血细胞中的表达变化；利用RNAi技术敲降tdch在对虾中的表达，研究其在对虾抗副溶血弧菌免疫中的作用。【结果与结论】克隆获得tdch的编码区全长，编码551氨基酸的蛋白，理论分子质量为63.37 ku，等电点为7.74;tdch除含有血蓝蛋白结构域以外，还含有酪氨酸酶Tyrosinase结构域；tdch在对虾肌肉、肝胰腺、鳃、胃、表皮、触角、心脏、肠、幽门盲囊、眼柄和神经索中均有表达，眼柄中表达最高，在幽门盲囊中表达最低。Poly(I:C)刺激后，tdch在血细胞中的表达显著升高，在鳃组织中表达变化不明显；其余5种免疫原刺激后，tdch在鳃和血细胞中的表达均显著升高，沉默tdch的表达可导致副溶血弧菌感染对虾存活率显著下降，表明tdch参与了抗副溶血弧菌免疫应答。

关键词(KeyWords)： 凡纳滨对虾;含酪氨酸酶结构域血蓝蛋白;免疫刺激;副溶血弧菌;免疫应答

Abstract:

Keywords:

基金项目(Foundation): 广东省基础与应用基础研究基金项目(2021A1515110900);; 中山大学中央高校基本科研业务费专项资金(22qntd2615)

作者(Author): 张锐,王子昂,汤俊宇,杨林蔚

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