糖蛋白G 基因在基因组P-M 位的插入重组表达对狂犬病毒Flury LEP 致病力的影响

Insertion of glycoprotein gene between P and M gene influences the pathogenicity of the rabies virus Flury LEP

摘要:

摘要: 【目的】研究狂犬病病毒Flury 鸡胚低代毒株(Flury LEP) 在基因组P-M 位增加糖蛋白基因(G基因)的重组表达对病毒致病力的影响。【方法】利用反向遗传操作技术，构建了P、M 基因之间额外插入G 基因的重组狂犬病病毒Flury LEP 株(rLEP-PGM)，并对重组病毒的生物学特性及对小鼠的致病性进行了初步研究。【结果】亲本株和重组病毒具有相似的生长特性，LEP和rLEP-PGM 在BHK-21 细胞的生长滴度分别为4×106 FFU/mL和2.5×106 FFU/mL，在小鼠神经母细

Abstract:

Abstract: ［Objective］To study the biological characteristics and pathogenicity of a recombinant rabies virus Flury LEP (low egg passage) that has two glycoprotein genes (Ggene)．［Methods］By using reverse genetics techniques，we constructed a recombinant virus Flury LEP that has an additional G gene between P and M gene (rLEP-PGM)．Then we studied the biological characteristics of the recombinant virus and its pathogenicity on mice．［Results］The in vitro growth characteristic of rLEP-PGM were similar to the LEP strain． Western blot analysis of glycoprotein expression showed that the glycoprotein expression level of rLEP-PGM was 1.5 times higher than LEP． The LD50 of rLEP-PGM and LEP was 3 FFU and 1 FFU by intracerebral injection． However，the LD50 of intramuscular injection was 4×104 Lg FFU and 3.2×105 Lg FFU，respectively．［Conclusion］Insertion of an additional G gene between P and M gene can significantly raise the expression level of glycoprotein and enhance the ability to invade central nervous system from peripheral sites．