文章摘要
孙玉苗,李富花,相建海.重组WSSV囊膜蛋白VP281和VP31的抗菌功能初步分析[J].水产学报,2012,36(12):1901~1909
重组WSSV囊膜蛋白VP281和VP31的抗菌功能初步分析
Preliminary judgment of the antibacterial functions of two recombinant WSSV envelope proteins VP281 and VP31
投稿时间:2011-10-21  修订日期:2012-04-15
DOI:DOI:10.3724/SP.J.1231.2012.27778
中文关键词: 白斑综合征病毒 VP281 VP31
英文关键词: white spot syndrome virus, WSSV, VP281, VP31
基金项目:国家重点基础研究发展计划
作者单位E-mail
孙玉苗 中国科学院海洋研究所 lanyueshusheng@163.com 
李富花 中国科学院海洋研究所  
相建海 中国科学院海洋研究所 jhxiang@qdio.ac.cn 
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中文摘要:
      摘要:由白斑综合征病毒(white spot syndrome virus, WSSV)引起的白斑综合症是危害虾产业最严重的传染性病毒病。病毒囊膜蛋白在与宿主细胞受体相互作用,启动病毒感染的发生或介导病毒的侵入方面起着重要作用。有关WSSV囊膜蛋白的研究较多,但目前尚未阐明各个囊膜蛋白的生理功能。本文尝试利用一种组成分泌型原核表达质粒pBTA1为表达载体,构建表达囊膜蛋白VP28、VP281或VP31的重组大肠杆菌菌株DH5α。组成型分泌rVP28和rVP281的重组菌构建成功,但含rVP281的重组菌与含rVP28的重组菌相比生长十分缓慢,推测rVP281的表达可能抑制大肠杆菌的生长。而组成型分泌rVP31的重组菌的构建未获成功。为弄清组成型分泌rVP31的重组菌未能成功构建的原因,使用pET-30a( )为表达载体,构建了表达rVP31包涵体的重组菌株BL21 (DE3) pLysS。将rVP31蛋白纯化并复性后,采用牛津杯法,检测rVP31蛋白的抗菌活性。以溶壁微球菌为受试菌,出现抑菌圈,表明rVP31对其具有抗菌作用。在动物病毒中发现具有抗菌作用的囊膜蛋白,可以增加有关WSSV的病毒学方面的知识。
英文摘要:
      Abstract: White spot syndrome caused by white spot syndrome virus (WSSV) is the most severe epidemic virosis to shrimp culture industry in the world, resulting in tremendous economic losses. Virus envelope proteins play important roles in interacting with host cells, initiating virus infection or mediating virus intrusion. Studies about WSSV envelope proteins are numerous, but their functions haven’t been elucidated. In this study, by using a prokaryotic constitutive secretory expression plasmid as the expression vector, the recombinant Escherichia coli strain DH5α that could constitutively secrete WSSV envelope protein VP28, VP281 or VP31 were tried to be constructed. Recombinant DH5α that could constitutively secrete rVP28 and rVP281 were constructed successfully and were respectively named DhpVP28 DhpVP281. DhpVP281 grew much slowly compared with DhpVP28 and we guessed that the expression of rVP281 probably inhibited the growth of E. coli. However, recombinant bacteria constitutively expressing VP31 can not be obtained. In order to find out why recombinant bacteria that constitutively secrete rVP31 can not be constructed, recombinant bacteria BL21 (DE3) pLysS were constructed with pET-30a( ) as the expression vector and rVP31 was obtained in the form of inclusion body. After renaturation, the antibacterial activity of rVP31 was tested. The result showed that rVP31 has the antibacterial activity against Micrococcus lysodeikticus. These findings, reported for the first time, may increase the virology knowledge of WSSV.
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