Nsp9基因在Marc-145细胞上对猪繁殖与呼吸综合征病毒复制的影响

doi:10.16431/j.cnki.1671-7236.2016.12.005

《中国畜牧兽医》 ›› 2016, Vol. 43 ›› Issue (12): 3114-3120.doi: 10.16431/j.cnki.1671-7236.2016.12.005

Nsp9基因在Marc-145细胞上对猪繁殖与呼吸综合征病毒复制的影响

赵孟孟^1,2, 冯松林², 王文佳³, 邢星⁴, 冯嘉萍², 张桂红²

1. 河南农业大学牧医工程学院, 郑州 450002;
2. 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, 广州 510642;
3. 河南牧业经济学院制药工程学院, 郑州 450046;
4. 广西钦州保税港区出入境检验检疫局, 钦州 535008

收稿日期:2016-02-16 出版日期:2016-12-20 发布日期:2016-12-22
通讯作者: 张桂红 E-mail:guihongzh@scau.edu.cn
作者简介:赵孟孟(1986-),男,河南汝州人,博士,研究方向:猪繁殖与呼吸综合征病毒的致病机理,E-mail:zhaomengmeng502@163.com
基金资助:
国家自然科学基金：猪繁殖与呼吸综合征病毒非结构蛋白Nsp9功能研究（31272564）；公益性行业（农业）科研专项经费（201203039）；国家生猪现代农业产业技术体系项目（CARS-36）；国家重点研发计划课题（2016YFD0500707）

Effect of Nsp9 Gene on Replication of PRRSV in Marc-145 Cells

ZHAO Meng-meng^1,2, FENG Song-lin², WANG Wen-jia³, XING Xing⁴, FENG Jia-ping², ZHANG Gui-hong²

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

Received:2016-02-16 Online:2016-12-20 Published:2016-12-22

摘要/Abstract

摘要：

试验旨在验证Nsp9基因是否影响猪繁殖与呼吸综合征病毒（porcine reproductive and respiratory syndrome virus，PRRSV）的复制。构建含有Nsp9全长基因的质粒pIRES2-EGFP-Nsp9，并转染到Marc-145细胞中，接毒之后分别通过实时荧光定量PCR和Western blotting方法测定PRRSV N蛋白在mRNA和蛋白水平的表达。结果显示，转染Nsp9基因的Marc-145细胞上PRRSV的N蛋白在mRNA水平上显著高于未转染Nsp9基因的对照组（P<0.05），是对照组的1.5倍，同时转染Nsp9基因后的Marc-145上PRRSV N蛋白水平也明显高于对照组。随着剂量的增加，N蛋白的表达量在mRNA和蛋白水平都有所增加。由以上结果初步可知，Nsp9基因可以在Marc-145细胞上促进PRRSV的复制，Nsp9基因与其复制密切相关。

关键词: 猪繁殖与呼吸综合征病毒; Nsp9基因; Marc-145细胞; 复制

Abstract:

In order to evaluate whether Nsp9 could enhance the replication of porcine reproductive and respiratory syndrome virus (PRRSV), pIRES2-EGFP-Nsp9 plasmid containing whole Nsp9 genome were transfected into Marc-145 cells,Real-time PCR and Western blotting were used to evaluate the expression of N protein after PRRSV was inoculated. The results showed that the level of N protein in the Nsp9 transfected cells was significantly higher than control group (P<0.05),which was 1.5 times higher. Meanwhile,the expression at protein level had the same trend as the mRNA level. When the dose of plasmid was increasing,the expression of N protein both at the mRNA and protein levels were increased. In conclusion,Nsp9 gene could enhance the replication of PRRSV in Marc-145 cells.

Key words: PRRSV; Nsp9 gene; Marc-145 cell; replication

中图分类号:

Q785

赵孟孟, 冯松林, 王文佳, 邢星, 冯嘉萍, 张桂红. Nsp9基因在Marc-145细胞上对猪繁殖与呼吸综合征病毒复制的影响[J]. 《中国畜牧兽医》, 2016, 43(12): 3114-3120.

ZHAO Meng-meng, FENG Song-lin, WANG Wen-jia, XING Xing, FENG Jia-ping, ZHANG Gui-hong. Effect of Nsp9 Gene on Replication of PRRSV in Marc-145 Cells[J]. , 2016, 43(12): 3114-3120.

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Nsp9基因在Marc-145细胞上对猪繁殖与呼吸综合征病毒复制的影响

Effect of Nsp9 Gene on Replication of PRRSV in Marc-145 Cells

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