李景芬;刘莉;曹访;

• 1:湖州师范学院生命科学学院

摘要(Abstract)：

[目的]克隆草鱼的PKR基因,为草鱼抗病毒基因研究奠定基础。[方法]依据GenBank上斑马鱼(AJ852023.1)和鲫鱼(AY293929.1)的PKR基因序列,利用Primer Premier 5.0软件设计了3对简并引物;采用100μg/ml PolyI:C体外分别处理草鱼肾细胞(Ctenopharyngodon indellus kidney cells,CIK)12、36、48 h,并提取处理后细胞的总RNA,逆转录后用降落PCR方法扩增这3个处理时间细胞中PKR基因。[结果]处理12 h时未扩增出PKR基因,处理36和48 h时都扩增到了PKR基因,并且表达量随处理时间的增加有所升高,扩增到的部分序列与鲫鱼和斑马鱼的该段序列同源性分别为100.00%和81.48%。[结论]试验成功获得了草鱼PKR基因的部分序列,PolyI:C高效诱导草鱼PKR蛋白表达将有助于开创治疗草鱼类病毒病的一种新思路。

关键词(KeyWords)： PolyI:C;PKR;草鱼;克隆;表达

Abstract:

Keywords:

基金项目(Foundation): 浙江省自然科学基金资助项目(Y3110432);; 湖州师范学院校级科研资助项目

作者(Author): 李景芬;刘莉;曹访;

Email:

DOI: 10.13989/j.cnki.0517-6611.2011.33.224

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