Han Mingyuan, Ke Hanzhong, Du Yijun, Zhang Qingzhan, Yoo Dongwan
Department of Pathobiology, University of Illinois at Urbana-Champaign, Urbana, IL, 61802, USA.
Department of Pediatrics and Communicable Diseases, University of Michigan Medical School, Ann Arbor, MI, 48109, USA.
Methods Mol Biol. 2017;1602:29-46. doi: 10.1007/978-1-4939-6964-7_3.
Porcine reproductive and respiratory syndrome (PRRS) is a reemerging swine disease, and has become economically the most significant disease in pork production worldwide. The causative agent is PRRS virus (PRRSV), which is a member virus of the family Arteriviridae. The PRRSV genome is a single-stranded positive-sense RNA and is infectious. Two strategies in the PRRSV reverse genetics system have been employed for reconstitution of progeny virus: RNA transfection and DNA transfection. The PRRSV reverse genetics has broadly been used for studies including protein structure-function relationship, foreign gene expression, vaccine development, virulence determinants, and viral pathogenesis. Herein, we describe the modification of the pFL12 "RNA launch" reverse genetic system to the CMV promoter-driven pXJ41-FL13 "DNA launch" system. The generation of progeny PRRSV using pXJ41-FL13 is further elucidated.
猪繁殖与呼吸综合征(PRRS)是一种再度出现的猪病,在全球猪肉生产中已成为经济上最重要的疾病。病原体是PRRS病毒(PRRSV),它是动脉炎病毒科的一种成员病毒。PRRSV基因组是单链正链RNA且具有感染性。PRRSV反向遗传系统中有两种策略被用于子代病毒的重建:RNA转染和DNA转染。PRRSV反向遗传学已广泛应用于包括蛋白质结构 - 功能关系、外源基因表达、疫苗开发、毒力决定因素和病毒发病机制等研究。在此,我们描述了将pFL12“RNA启动”反向遗传系统改造为CMV启动子驱动的pXJ41 - FL13“DNA启动”系统。进一步阐明了使用pXJ41 - FL13产生子代PRRSV的过程。