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VP31的序列，设计并合成特异引物，PCR扩增得到VP31基因，大小为783bp。通过引物两端的BamHⅠ和EcoRⅠ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31kD大小相吻合的蛋白带。用Ni^2＋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。[著者文摘]
文章出处:
《上海水产大学学报》-2008年1期 -47-51页
分 类 号:
文献标识码:
A
文章编号:
1004-7271（2008）01-0047-05
Relationship of white spot syndrome virus envelope protein VP31 with precociousness of WSSV
WANG Yong-jie, PANG Ting-shuang, LI Yan-he, SHE Lei （ Institute of Fisheries, Anhui Academy of Agricultural Sciences, Hefei 230031, China）
Abstract:
White spot syndrome virus （WSSV） is a devastating viraI pathogen of cultured shrimp worIdwide. In this paper, two primers were designed according to the sequences of envelope genes, VP31 of white spot syndrome virus（WSSV） in the GenBank. The DNA fragment 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 VP31 purified by Ni^2＋ column chromatography was prepared for immunizing New Zealand rabbit. The VP31 antiserum was used to neutralize the WSSV infection on crayfish by intramuscular injection. A neutralization assay with this antibody demonstrated that envelope protein VP31 is involved in WSSV infection and that VP31 might play a key role during this process.[著者文摘]
Key words:
crayfish; white spot syndrome virus; envelope protein VP31 ; in vivo neutralization
基金资助:
国家重点基础研究973发展计划（2005CB121000）;安徽省农科院院长创新基金（2006-76）
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