对虾白斑综合征病毒的细胞因子受体基因的分析与表达  

Sequence Analysis, Cloning and Expression of a Putative Cytokine Receptor Gene of White spot syndrome virus

作　　者：解云礼[1] 黄茹[1] 石正丽[1] 

机构地区：[1]中国科学院武汉病毒研究所分子病毒学实验室无脊椎动物病毒学联合开放实验室,湖北武汉430071

出　　处：《中国病毒学》2003年第4期362-366,共5页Virologica Sinica

基　　金：中国科学院生命科学与生物技术领域2001年知识创新工程重要方向项目(KSCX2-SW-302-2);863海洋生物技术课题(2001AA620201)

摘　　要：在对虾白斑综合征病毒(White spot syndrome virus,WSSV)的基因组中发现一个具有细胞因子受体特征的开放阅读框,该阅读框全长2022个核苷酸,编码674个氨基酸,蛋白质理论分子量为76kDa。该基因含有真核生物细胞因子gp130受体特征序列。为了研究该基因的功能,采用PCR方法从病毒基因组中扩增出基因片段,克隆到pGEM-T Easy载体中,经BamH Ⅰ和Sal Ⅰ双酶切后插入pET28b表达载体中。重组质粒转化到大肠杆菌BL21中,IPTG诱导后,经SDS-PAGE电泳表明在76kDa处有目的蛋白表达。用冰浴超声波对诱导后的菌液进行处理以获得初步纯化的蛋白,作为抗原人工免疫实验兔子以获得含特异性抗体的抗血清。该基因的表达成功,为其功能的进一步深入研究奠定了基础。One ORF of White spot syndrome virus (WSSV) genome has homology with the cytokine receptor. This ORF contains 2022 nucleotides and codes for 674 amino acids with a theoretical molecular weight of 76 kDa. This putative gene contains one conserved motif of eukaryotic cytokine receptor gp130. To investigate the function of this protein, the gene was amplified from the genome of WSSV by polymerase chain reaction (PCR), then cloned into pET28b and expressed in E.coli. The expressed fusion protein tagged with six consecutive histidines was 76kDa after SDS-PAGE. The purified fusion protein was used as antigen to immunize rabbit for further gene function study.

关 键 词：对虾白斑综合征病毒 细胞因子受体 基因 表达 

分 类 号：S945.4[农业科学—水产养殖]