对虾白斑综合症病毒结构蛋白质VP12的研究

Study of white spot syndrome virus structural protein VP12

VP12是对虾白斑综合症病毒的一种结构蛋白,该蛋白质由病毒基因wsv009编码,分子量为10.9kDa.转录时相分析表明wsv009属于早期基因.本研究通过PCR扩增获得了该全长基因,成功构建了重组质粒pET-His-vp12.将该质粒转化到E.coliBL21中表达并纯化了重组VP12蛋白,用该重组蛋白制备了特异性鼠抗血清.纯化的病毒粒子经0.5%Triton X-100处理后,VP12被发现存在于病毒WSSV的囊膜部分,Western blot杂交实验和免疫电镜实验也证实该结果.对VP12的深入研究将有助于了解病毒的感染和包装的机制,为病毒的诊断和控制提供科学依据.

The characterization of VP12 we described here is one of the low-abundance structural proteins of WSSV.It was a 10.9kDa envelop protein from WSSV,being coded by ORF wsv009.The transcriptional analysis shows that wsv009 was an early gene.In this study,the full length of wsv009 gene was cloned into the plasmid pET-His to generate the recombinant plasmid pET-His-vp12.The pET-His-vp12 was then transformed into E.coliBL21 to express and purify the recombinant VP12.After that,using the purified recombinant VP12,the antiserum was produced.Meanwhile,the gene wsv009 was expressed in Escherichia coli and the recombinant VP12 was purified,then it was used to prepare specific antibody.When purified virions were treated with 0.5％ Triton X-100 detergent,VP12 was found to present exclusively in the envelope fraction,which was also confirmed by western blot analysis and immunoelectron microscopy.The result indicated that VP12 behaved as an envelope protein.The further study of VP12 may facilitate a better understanding of the molecular mechanism underlying WSSV package and may be helpful for the diagnosis and control of virus infection.