PRRSV强、弱毒株Nsp9基因对PRRSV复制的影响

发布时间：2018-04-18 08:28

本文选题：猪繁殖与呼吸综合征病毒 + Nsp基因　；参考：《华北农学报》2017年01期

【摘要】：为了探索猪繁殖与呼吸综合征病毒(PRRSV)强、弱毒株的Nsp9基因对PRRSV复制的影响,构建含有PRRSV XH-GD强毒株、CH-1R弱毒株Nsp9全长基因的质粒p IRes2-EGFP-Nsp9-XH-GD、p IRes2-EGFP-Nsp9-CH-1R,并将重组质粒分别转染到Marc-145细胞中,以MOI=1的剂量接种PRRSV XH-GD毒株,通过TCID50试验测定病毒的滴度,采用荧光定量PCR和Western Blot方法来测定病毒的N蛋白表达情况。结果表明,转染弱毒株CH-1R Nsp9基因的Marc-145细胞中,PRRSV的N蛋白在mRNA水平、蛋白质水平上的表达量均高于转染强毒株XH-GD组,且病毒的滴度也显著高于转染强毒株XH-GD组。综上,在Marc-145细胞中,弱毒株CH-1R Nsp9基因对PRRSV复制的促进作用优于强毒株XH-GD。
[Abstract]:In order to explore the effect of Nsp9 gene of porcine reproductive and respiratory syndrome virus (PRRSVV) and attenuated strain on PRRSV replication, the plasmid pIRes2-EGFP-Nsp9-XH-GDN pRes2-EGFP-Nsp9-CH-1R containing the full-length Nsp9 gene of PRRSV XH-GD virulent strain was constructed, and the recombinant plasmid was transfected into Marc-145 cells.The PRRSV XH-GD strain was inoculated with the dose of MOI=1, the titer of the virus was determined by TCID50 test, and the expression of N protein was detected by fluorescence quantitative PCR and Western Blot.The results showed that the expression of N protein in Marc-145 cells transfected with attenuated CH-1R Nsp9 gene was significantly higher than that in XH-GD group at the mRNA level, and the titer of the virus was significantly higher than that in the XH-GD group.In conclusion, in Marc-145 cells, the promotion of PRRSV replication by CH-1R Nsp9 gene of attenuated strain is superior to that of XH-GD strain.
【作者单位】：华南农业大学兽医学院国家生猪种业工程技术研究中心广东省动物源性人兽共患病预防与控制重点实验室;河南农业大学牧医工程学院;河南省农业科学院农业经济与信息研究所;河南牧业经济学院制药工程学院;广西钦州保税港区出入境检验检疫局;
【基金】：国家自然科学基金项目(31272564) 公益性行业(农业)科研专项(201203039) 国家生猪现代农业产业技术体系项目(CARS-36)
【分类号】：S852.651

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