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

XIE Yun—li; HUANG Ru; SHI Zheng—li

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August 2003

XIE Yun—li, HUANG Ru and SHI Zheng—li. Sequence Analysis，Cloning and Expression of a Putative Cytokine Receptor Gene of White spot syndrome virus[J]. Virologica Sinica, 2003, 18(4): 362-366.

Citation: XIE Yun—li, HUANG Ru, SHI Zheng—li. Sequence Analysis，Cloning and Expression of a Putative Cytokine Receptor Gene of White spot syndrome virus .VIROLOGICA SINICA, 2003, 18(4) : 362-366.

对虾白斑综合征病毒的细胞因子受体基因的分析与表达

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

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gpl30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。
- 对虾白斑综合征病毒(WSSV)
- , 基因表达
- , 细胞因子受体

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

1.
Key laboratory ofMolecular Virology,Joint—Laboratory ofInvertebrate Virology,Wuhan Institute of Virology,ChineseAcademy ofSciences,Wuhan 430071，China

Abstract

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．Th is putative gene contains one conserved motif of eukaryotic cytokine receptor gpl30．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．Th e expressed fusion protein tagged with six consecutive histidines was 76kDa after SDS—PAGE．Th e purified fusion protein was used as antigen to immunize rabbit for further gene function study．
- White spot syndrome virus(WSSV)
- , Gene expression
- , Cytokine receptor

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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

1. Key laboratory ofMolecular Virology,Joint—Laboratory ofInvertebrate Virology,Wuhan Institute of Virology,ChineseAcademy ofSciences,Wuhan 430071，China

Abstract: 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．Th is putative gene contains one conserved motif of eukaryotic cytokine receptor gpl30．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．Th e expressed fusion protein tagged with six consecutive histidines was 76kDa after SDS—PAGE．Th e purified fusion protein was used as antigen to immunize rabbit for further gene function study．