靶向cofilin2基因RNAi对鸭病毒性肠炎病毒增殖的影响

doi:10.11843/j.issn.0366-6964.2022.03.032

畜牧兽医学报 ›› 2022, Vol. 53 ›› Issue (3): 983-989.doi: 10.11843/j.issn.0366-6964.2022.03.032

靶向cofilin2基因RNAi对鸭病毒性肠炎病毒增殖的影响

王微¹, 华敏¹, 张芸², 李涛³, 张黔东¹, 袁阳¹, 程振涛^1,4, 文明^1,4*

1. 贵州大学动物科学学院, 贵阳 550025;
2. 贵州省畜禽遗传资源管理站, 贵阳 550025;
3. 贵州省动物预防控制中心, 贵阳 550025;
4. 贵州省动物生物制品工程技术研究中心, 贵阳 550025

收稿日期:2021-05-28 出版日期:2022-03-23 发布日期:2022-03-31
通讯作者: 文明,主要从事病原分子生物学与免疫学研究,E-mail:as.mwen@gzu.edu.cn
作者简介:王微(1998-),女,贵州石阡人,硕士生,主要从事动物疾病防控研究,E-mail:2431196391@qq.com
基金资助:
国家自然科学基金项目(31260607;31560703);贵州省百层次创新型人才项目(黔科合人才[2016]4009号);贵州省科技平台及人才团队计划项目(黔科合平台人才[2018]5253号)

Effect of RNAi Targeting cofilin2 Gene on Duck Enteritis Virus Proliferation

WANG Wei¹, HUA Min¹, ZHANG Yun², LI Tao³, ZHANG Qiandong¹, YUAN Yang¹, CHENG Zhentao^1,4, WEN Ming^1,4*

1. College of Animal Sciences, Guizhou University, Guiyang 550025, China;
2. Guizhou Province Livestock and Poultry Resources Genetic Management Station, Guiyang 550025, China;
3. Animal Prevention and Control Center of Guizhou, Guiyang 550025, China;
4. Guizhou Animal Biological Products Engineering and Technology Research Center, Guiyang 550025, China

Received:2021-05-28 Online:2022-03-23 Published:2022-03-31

摘要/Abstract

摘要： 旨在探讨cofilin2基因的表达在鸭病毒性肠炎病毒(DEV)增殖过程中的影响。本研究通过已获得的鸭源cofilin2全基因序列(GenBank登录号:MF434775),设计出4对shRNA序列,插入载体中,并转染到鸭胚成纤维细胞中,经FQ-PCR方法和荧光显微镜观察筛选出具有较高干扰效率的shRNA序列,用干扰效率较高的shRNA序列转染到鸭胚成纤维细胞中抑制cofilin2基因的表达,再检测cofilin2在DEV复制过程中表达情况。结果显示:4对shRNA干扰质粒经转染至鸭胚成纤维细胞后,经荧光显微镜观察,构建的4对shRNA干扰质粒均成功转染并得到表达;4对干扰质粒沉默率分别为4.96%±0.78%、41.12%±2.94%、17.55%±2.07%、54.02%±1.73%,以cofilin2-86的沉默效率最高;cofilin2-86转染到鸭胚成纤维细胞,经DEV感染不同时间,其DEV核酸表达量明显下降。综上所述,靶向cofilin2基因RNA干扰在DEV复制和增殖过程发挥重要作用。

关键词: 鸭病毒性肠炎病毒, DEV, RNA干扰, cofilin2

Abstract: This study aimed to investigate the effect of cofilin2 gene expression on the proliferation of duck enteritis virus (DEV). The complete gene sequence of cofilin2 from duck (GenBank:MF434775),four pairs of shRNA sequences were designed,inserted into vectors and transfected into duck embryo fibroblasts. shRNA sequences with high interference efficiency were screened by FQ-PCR and fluorescence microscope observation. The shRNA sequences with high interference efficiency were transfected into duck embryo fibroblasts to inhibit cofilin2 gene expression. Then check how cofilin2 is expressed during DEV replication. Result were as follows:Four pairs of shRNA interfering plasmids were successfully transfected into duck embryo fibroblasts and expressed by fluorescence microscopy. The silences were 4.96%±0.78%, 41.12%±2.94%, 17.55%±2.07%, 54.02%±1.73%, respectively, and cofilin2-86 was the most efficient. After cofilin2-86 transfection into duck embryo fibroblasts and DEV infection, the expression of DEV nucleic acid decreased significantly at different time. In conclusion, RNA interference targeting cofilin2 gene plays an important role in DEV replication and proliferation.

Key words: duck enteritis virus, DEV, RNA interference, cofilin2

中图分类号:

S852.659.1

王微, 华敏, 张芸, 李涛, 张黔东, 袁阳, 程振涛, 文明. 靶向cofilin2基因RNAi对鸭病毒性肠炎病毒增殖的影响[J]. 畜牧兽医学报, 2022, 53(3): 983-989.

WANG Wei, HUA Min, ZHANG Yun, LI Tao, ZHANG Qiandong, YUAN Yang, CHENG Zhentao, WEN Ming. Effect of RNAi Targeting cofilin2 Gene on Duck Enteritis Virus Proliferation[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(3): 983-989.

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靶向cofilin2基因RNAi对鸭病毒性肠炎病毒增殖的影响

Effect of RNAi Targeting cofilin2 Gene on Duck Enteritis Virus Proliferation

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