Expression and functional analysis of Brucella outer membrane protein 25 in recombinant goat pox virus

布鲁氏杆菌外膜蛋白25在重组山羊痘病毒中的表达及功能分析

阅读:7
作者:Zhihua Sun, Laizhen Liu, Hui Zhang, Youwen Li, Feng Wei, Zhiqiang Li, Pengyan Wang, Qiang Fu, Yan Ren, Yu Zhang, Zhiru Guo, Chuangfu Chen

Abstract

The Capripoxvirus (CaPV) has a large double‑stranded DNA genome and a restricted host‑range. At present, it is being investigated as an ideal vaccine vector. In the present study, a novel recombinant goat pox virus (rGTPV) was constructed to express Brucella outer membrane protein (OMP)25, and was validated by in vitro and in vivo immunization assays. A novel rGTPV vector was created, in which the thymidine kinase gene was used as a flanking sequence, I1L was inserted as a promoter element to enhance Brucella OMP25 expression, and p7.5 as another promoter element was used to regulate guanine phosphoribosyl‑transferase as a selection maker. The rGTPV vector was transfected into sheep fetal fibroblast/lamb testis cells pre‑infected with GTPV G14‑STV44‑55 to recombine. Brucella OMP25 protein was expressed in cells by rGTPV, and activated immune reactivity to Brucella OMP25 protein, as detected by western blotting. Furthermore, rGTPV elicited, anti‑Brucella‑specific immunoglobulin G responses, as determined by ELISA. Mice vaccinated with rGTPV did not exhibit pathology alterations in the kidney and liver. These results suggested that the novel rGTPV was able to efficiently drive Brucella OMP25 protein expression and activate immune reactivity, and may have applications in CaPV live vector vaccines and associated research.

特别声明

1、本页面内容包含部分的内容是基于公开信息的合理引用;引用内容仅为补充信息,不代表本站立场。

2、若认为本页面引用内容涉及侵权,请及时与本站联系,我们将第一时间处理。

3、其他媒体/个人如需使用本页面原创内容,需注明“来源:[生知库]”并获得授权;使用引用内容的,需自行联系原作者获得许可。

4、投稿及合作请联系:info@biocloudy.com。