PRRSV强、弱毒株Nsp9基因对PRRSV复制的影响 |
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引用本文: | 赵孟孟,冯丽丽,冯松林,马红芳,张二芹,王文佳,邢星,张雅,冯嘉萍,张桂红.PRRSV强、弱毒株Nsp9基因对PRRSV复制的影响[J].华北农学报,2017,32(1). |
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作者姓名: | 赵孟孟 冯丽丽 冯松林 马红芳 张二芹 王文佳 邢星 张雅 冯嘉萍 张桂红 |
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作者单位: | 1. 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,广东广州510642;河南农业大学牧医工程学院,河南郑州450002;2. 河南省农业科学院农业经济与信息研究所,河南郑州,450002;3. 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,广东广州510642;4. 河南农业大学牧医工程学院,河南郑州,450002;5. 河南牧业经济学院制药工程学院,河南郑州,450046;6. 广西钦州保税港区出入境检验检疫局,广西钦州,535008 |
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基金项目: | 国家自然科学基金项目,公益性行业(农业)科研专项,国家生猪现代农业产业技术体系项目 |
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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。
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关 键 词: | 猪繁殖与呼吸综合征病毒 Nsp9基因 复制 病毒滴度 |
Influence of Nsp9 Between Low Virulent and Virulent Strains to Replication of PRRSV in Marc-145 Cells |
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Abstract: | In order to verify the difference of gene Nsp9 between virulent strain and low virulent strain whether could enchance the replication of PRRSV,plasmids pIRes2-EGFP-Nsp9-XH-GD and pIRes2-EGFP-Nsp9-CH-1R containing whole Nsp9 genome were transfected into Marc-145 cells,qPCR and Western Blot were conducted to e valuate its titer and expression after PRRSV was inoculated,the results showed that the level of N protein in the cells that were transfected with low virulent strain CH-1R Nsp9 gene was higher than the control group-virulent strain XH-GD in the mRNA level.Meanwhile,on the protein level,it was the same as the mRNA level,and virus titer was also higher than virulent strain.It demonstrated that Nsp9 gene of low virulent strain CH-1R in the Marc145 cells was more beneficial to the replication of PRRSV than that of virulent strain XH-GD. |
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Keywords: | PRRSV Nsp9 Replication Viraltiter |
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