Gromadzka Beata, Szewczyk Bogusław, Konopa Grazyna, Fitzner Andrzej, Kesy Andrzej
Department of Molecular Virology, Intercollegiate Faculty of Biotechnology, University of Gdańsk and Medical University of Gdańsk, Gdańsk, Poland.
Acta Biochim Pol. 2006;53(2):371-6. Epub 2006 May 30.
Rabbit hemorrhagic disease virus (RHDV) which causes a highly contagious disease of wild and domestic rabbits belongs to the family Caliciviridae. It is a small, positive single-stranded RNA virus with a genome of 7.5 kb and has a diameter of approximately 40 nm. In negatively stained electron micrographs the virus shows typical calicivirus morphology with regularly arranged cup-shaped structures on the surface. It is a major pathogen of rabbits in many countries. Vp60 - a coat protein of molecular mass around 60 kDa is the major antigen of RHDV. It is present as 90 dimeric units per virion particle. We have expressed VP60 gene in the baculovirus system with the aim to use it as a potential vaccine against RHDV and a diagnostic reagent in immunological tests. cDNA of the vp60 gene of strain SGM, was cloned into a baculovirus transfer vector as full-length gene, as well as truncated gene lacking 600 5'-terminal nucleotides. The sequence of SGM VP60 differed markedly from that of the reference strain. Full-length recombinant VP60 protein from the SGM strain self-assembled to form virus-like particles (VLPs). These particles observed by electron microscopy were morphologically similar to native virions and were able to agglutinate human group 0 erythrocytes. After immunization the recombinant particles induced RHDV-specific antibodies in rabbits and guinea pigs. Rabbits immunized with the VLPs were fully protected against challenge with a virulent RHDV.
兔出血症病毒（RHDV）可引发家兔和野兔的一种高度传染性疾病，属于杯状病毒科。它是一种小型的正链单链RNA病毒，基因组为7.5 kb，直径约为40纳米。在负染电子显微镜照片中，该病毒呈现典型的杯状病毒形态，表面有规则排列的杯状结构。它是许多国家家兔的主要病原体。Vp60——一种分子量约为60 kDa的衣壳蛋白，是RHDV的主要抗原。每个病毒粒子中以90个二聚体单位形式存在。我们在杆状病毒系统中表达了VP60基因，目的是将其用作抗RHDV的潜在疫苗以及免疫检测中的诊断试剂。将SGM株vp60基因的cDNA作为全长基因以及缺失5'端600个核苷酸的截短基因克隆到杆状病毒转移载体中。SGM VP60的序列与参考株明显不同。来自SGM株的全长重组VP60蛋白自组装形成病毒样颗粒（VLP）。通过电子显微镜观察到的这些颗粒在形态上与天然病毒粒子相似，并且能够凝集人0型红细胞。免疫后，重组颗粒在兔和豚鼠体内诱导产生RHDV特异性抗体。用VLP免疫的兔对强毒RHDV攻击具有完全的抵抗力。
