High-level expression of rabies virus glycoprotein with the RNA-based Semliki Forest Virus expression vector

被引:17
作者
Benmaamar, Rim [2 ,3 ]
Astray, Renato Mancini [1 ]
Wagner, Renaud [2 ]
Pereira, Carlos Augusto [1 ]
机构
[1] Inst Butantan, Lab Imunol Viral, BR-05503900 Sao Paulo, Brazil
[2] CNRS, UMR7175, Dpt Recepteurs & Prot Membranaires, Illkirch Graffenstaden, France
[3] Bioxtal, CH-1006 Epalinges, Switzerland
基金
巴西圣保罗研究基金会;
关键词
Rabies virus; Rabies virus glycoprotein (RVGP); Semliki Forest Virus (SFV); MELANOGASTER CELLS; RECEPTOR; LYSSAVIRUS; SYSTEM; PARTICLES; VERSATILE; PROTEIN; BURDEN; GENE;
D O I
10.1016/j.jbiotec.2008.12.009
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Rabies is to this date one of the most important death causing zoonotic viral diseases, with 98% of deaths reported in developing countries, where access to modern vaccines and tools for efficient diagnostic remain Unaffordable. in this paper, we describe a newly engineered RNA-based rabies virus glycoprotein (RVGP) expression vector based on the Semliki Forest Virus (SFV) system. A recombinant SFV carrying an RNA coding for RVGP (SFV-RVGP) was constructed and the RVGP expression was evaluated in animal cell cultures. The mRNA coding for RVGP and the RVGP itself were assessed by qPCR, Western-blotting, confocal microscopy, flow cytometry and ELISA. Moreover, SFV-RVGP was proven to be highly efficient in expressing the functionally trimeric RVGP, SFV-RVGP is easy to produce and efficient in different cell lines, making it an interesting candidate for efficient and functional viral glycoprotein expression. (c) 2008 Elsevier B.V. All rights reserved.
引用
收藏
页码:283 / 290
页数:8
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