PRRSV强、弱毒株<em>Nsp9</em>基因对PRRSV复制的影响

doi:10.7668/hbnxb.2017.01.003

华北农学报 ›› 2017, Vol. 32 ›› Issue (1): 15-20. doi: 10.7668/hbnxb.2017.01.003

所属专题： 生物技术；

PRRSV强、弱毒株Nsp9基因对PRRSV复制的影响

赵孟孟^1,2, 冯丽丽³, 冯松林¹, 马红芳², 张二芹², 王文佳⁴, 邢星⁵, 张雅², 冯嘉萍¹, 张桂红¹

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;
2. 河南农业大学牧医工程学院, 河南郑州 450002;
3. 河南省农业科学院农业经济与信息研究所, 河南郑州 450002;
4. 河南牧业经济学院制药工程学院, 河南郑州 450046;
5. 广西钦州保税港区出入境检验检疫局, 广西钦州 535008

收稿日期:2017-03-20 出版日期:2017-02-28
通讯作者: 张桂红(1968-),女,黑龙江哈尔滨人,教授,博士,主要从事动物传染病研究。
作者简介:赵孟孟(1986-),男,河南汝州人,在读博士,主要从事猪繁殖与呼吸综合征病毒的致病机理研究。
基金资助:
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）

Influence of Nsp9 Between Low Virulent and Virulent Strains to Replication of PRRSV in Marc-145 Cells

ZHAO Mengmeng^1,2, FENG Lili³, FENG Songlin¹, MA Hongfang², ZHANG Erqin², WANG Wenjia⁴, XING Xing⁵, ZHANG Ya², FENG Jiaping¹, ZHANG Guihong¹

1. College of Veterinary Medicine, South China Agricultural University, National Engineering Research Center for Breeding Swine Industry, Key Laboratory of Zoonosis Prevention and Control of Guangdong Province, Guangzhou 510642, China;
2. College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China;
3. Institute of Agricultural Economics and Information, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China;
4. School of Pharmaceutical Engineering, Henan University of Animal Husbandry and Economy, Zhengzhou 450046, China;
5. Guangxi Qinzhou Free Trade Port Area Entry-exit Inspection and Quarantine, Qinzhou 535008, China

Received:2017-03-20 Published:2017-02-28

摘要/Abstract

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pIRes2-EGFP-Nsp9-XH-GD、pIRes2-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。

关键词: 猪繁殖与呼吸综合征病毒, Nsp9基因, 复制, 病毒滴度

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 evaluate 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 Marc-145 cells was more beneficial to the replication of PRRSV than that of virulent strain XH-GD.

Key words: PRRSV, Nsp9, Replication, Viraltiter

中图分类号:

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): 15-20. doi: 10.7668/hbnxb.2017.01.003.

ZHAO Mengmeng, FENG Lili, FENG Songlin, MA Hongfang, ZHANG Erqin, WANG Wenjia, XING Xing, ZHANG Ya, FENG Jiaping, ZHANG Guihong. Influence of Nsp9 Between Low Virulent and Virulent Strains to Replication of PRRSV in Marc-145 Cells[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(1): 15-20. doi: 10.7668/hbnxb.2017.01.003.

http://www.hbnxb.net/CN/Y2017/V32/I1/15

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