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在大肠杆菌中表达的兔出血症病毒衣壳蛋白病毒样颗粒的自组装及其在兔体内的免疫原性。

Self-assembly of virus-like particles of rabbit hemorrhagic disease virus capsid protein expressed in Escherichia coli and their immunogenicity in rabbits.

作者信息

Guo Huimin, Zhu Jie, Tan Yonggui, Li Chuanfeng, Chen Zongyan, Sun Shiqi, Liu Guangqing

机构信息

Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 200241, Shanghai, China.

Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 730046, Gansu, China.

出版信息

Antiviral Res. 2016 Jul;131:85-91. doi: 10.1016/j.antiviral.2016.04.011. Epub 2016 Apr 23.

Abstract

In this study, virus-like particles (VLPs) derived from rabbit hemorrhagic disease virus (RHDV) were evaluated for the development of a vaccine against RHDV infection. The VP60 gene was cloned and inserted into a pSMK expression vector containing a small ubiquitin-like modifier (SUMO) tag that can promote the soluble expression of heterologous proteins in Escherichia coli cells. After expression and purification of His-SUMO-VP60 and cleavage of the SUMO tag, we found that the RHDV VP60 protein had self-assembled into VLPs with a similar shape and smaller size compared with authentic RHDV capsid. Next, the antigenicity and immunogenicity of the VLPs were examined. The results showed that RHDV-specific responses were clearly induced in rabbits and that all rabbits in the VLP group survived while those in the negative control group died within 72 h post-infection. These results suggest that VLP-based RHDV could be a promising RHDV vaccine candidate.

摘要

在本研究中,对源自兔出血症病毒(RHDV)的病毒样颗粒(VLPs)进行了评估,以开发一种针对RHDV感染的疫苗。克隆了VP60基因,并将其插入到一个含有小泛素样修饰物(SUMO)标签的pSMK表达载体中,该标签可促进异源蛋白在大肠杆菌细胞中的可溶性表达。在表达并纯化His-SUMO-VP60并切割SUMO标签后,我们发现RHDV VP60蛋白已自组装成VLP,其形状与天然RHDV衣壳相似,但尺寸较小。接下来,检测了VLP的抗原性和免疫原性。结果表明,在兔中明显诱导了RHDV特异性反应,VLP组的所有兔子均存活,而阴性对照组的兔子在感染后72小时内死亡。这些结果表明,基于VLP的RHDV可能是一种有前景的RHDV疫苗候选物。

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