对虾白斑综合症病毒类锌指蛋白基因(wsv063、wsv069和wsv477)的表达和纯化  

Expression of Three Putative Zinc Finger Protein Genes wsv063,wsv069 and wsv477 in White Spot Syndrome Virus(WSSV) Genome and Their Products Purification

作　　者：朱艳冰[1] 杨丰[2] 

机构地区：[1]厦门大学生命科学学院 [2]国家海洋局第三海洋研究所,福建厦门361005

出　　处：《厦门大学学报（自然科学版）》2005年第5期697-700,共4页Journal of Xiamen University：Natural Science

基　　金：国家自然科学基金(30330470);福建省自然科学基金(2003F001)资助

摘　　要：从对虾白斑综合症病毒基因组的预测开放阅读框中选取3个编码类锌指蛋白的基因wsv063、wsv069和wsv477,将它们克隆到pET-GST表达载体,以E.coliBL21(DE3)为宿主菌,成功表达了GST融合的目的蛋白.表达的GST-WSV063和GST-WSV477在菌体里主要以包涵体形式存在,GST-WSV069在菌体里可溶性形式和包涵体形式几乎等量存在.采用glutathione sepharose 4B或Ni-NTA agarose亲和层析对目的蛋白进行了纯化.以上3种病毒蛋白的表达和纯化为进一步研究它们的功能奠定了基础.Three open reading frames （termed wsv063,wsv069 and wsv477） of white spot syndrome virus are predicted to encode putative zinc finger proteins. They were cloned and inserted into pET-GST vector and expressed in E. coli BL21 （DE3） ,and then the glutathione S-transferase （GST） fusion proteins with high yield were obtained. It was found that GST-WSV063 and GST-WSV477 fusion proteins existed mainly in inclusion bodies of E. coli and GST-WSV069 in soluble forms and inclusion bodies both. Subsequently,the fusion proteins were purified using glutathione sepharose 4B or Ni-NTA agarose affinity chromatography. The expression of wsv063,wsv069 and wsv477 genes and theirs products purification set a satisfactory foundation for their function researches.

关 键 词：对虾白斑综合症病毒wsv63 wsv69和wsv477基因 类锌指蛋白 表达和纯化 

分 类 号：S852.65[农业科学—基础兽医学]