白斑综合征病毒囊膜蛋白VP31与该病毒侵染的相关性.pdfVIP

第l7卷第1期 上 海 水 产 大 学 学 报 Vo1．17．No．1 2008年1月 JOURNAL OF SHANGHAI FISHERIES UNIVERSITY Jan．，2008 文章编号：1004—7271(2008)01—0047—05 白斑综合征病毒囊膜蛋白VP31与 该病毒侵染的相关性 王永杰，潘庭双，李艳和，佘 磊 (安徽省农业科学院水产研究所，安徽合肥 230031) 摘 要：白斑综合征病毒(WSSV)是一种已给世界养虾业造成了严重危害的病原体。根据GenBank上公布的 WSSV囊膜蛋白基因VP3l的序列，设计并合成特异引物，PCR扩增得到VP3l基因，大小为783 bp。通过引物 两端的BamH I和EcoR I酶切位点连接该基因，然后将其构建到家蚕杆状病毒表达载体pFAST BAC HTB上。 重组的家蚕杆状病毒转染5龄家蚕幼体内表达，表达产物经SDS—PAGE检测，显示与预期3l kD大小相吻合 的蛋白带。用Ni 柱纯化该蛋白免疫新西兰大白兔制备抗体，肌肉注射克氏鳌虾进行活体中和病毒试验。 中和试验结果显示，囊膜蛋白VP31与白斑综合征病毒的侵染性相关，对病毒的侵染性可能起着重要作用。 关键词：克氏鳌虾；白斑综合征病毒；囊膜蛋白VP3l；体内中和试验 中图分类号：S 9l7 文献标识码：A Relationship of white spot syndrome virus envelope protein VP3 1 with prec0ci0usness of WSSV WANG Yong-jie，PANG Ting-shuang，LI Yan-he，SHE Lei (Institute of Fisheries，Anhui Academy of Agricultural Sciences，Hefei 23003 1，China) Abstract：White spot syndrome virus(WSSV)is a devastating viraI pathogen of cultured shrimp worldwide．In this paper，two primers were designed according to the sequences of envelope genes，VP3 1 of white spot syndrome virus(WSSV)in the GenBank．The DNA~agment about 783 bp amplified by PCR were linked to the BamH I and EcoR I restriction endonuclease site，then cloned into pFAST BAC HTB and expressed in the 5th instar larvae of silkworm．The molecular weight of the engineered protein was about 31 kD，which was identified by SDS—PAGE analysis． Antiserum of envelope protein VP3 1 purified by Ni column chromatography was prepared for immunizing New Zealand rabbit．The VP3 1 antiserum was used to neutralize the WSSV infection on crayfish by intramuscular injection．A neutralization assay with this antibody demonstrated that envelope protein VP3 l is involved in WSSV infection and that