VP3 G–H环中氨基酸突变对O型口蹄疫病毒生物学特性的影响 Effect of amino acid mutation(s) in the G-H loop of VP3 on biological characteristics of foot-and-mouth disease virus serotype O期刊界 All Journals 搜尽天下杂志传播学术成果专业期刊搜索期刊信息化学术搜索

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VP3 G–H环中氨基酸突变对O型口蹄疫病毒生物学特性的影响

引用本文：	陈冬冬,白兴文,宫晓华,包慧芳,李平花,李冬,付元芳,白启峰,袁红,孙普,马雪青,曹轶梅,陈应理,卢曾军,刘在新.VP3 G–H环中氨基酸突变对O型口蹄疫病毒生物学特性的影响[J].微生物学报,2019,59(7):1285-1294.

作者姓名：	陈冬冬白兴文宫晓华包慧芳李平花李冬付元芳白启峰袁红孙普马雪青曹轶梅陈应理卢曾军刘在新

作者单位：	中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, OIE/国家口蹄疫参考实验室, 甘肃兰州 730046,中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, OIE/国家口蹄疫参考实验室, 甘肃兰州 730046,中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, OIE/国家口蹄疫参考实验室, 甘肃兰州 730046,中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, OIE/国家口蹄疫参考实验室, 甘肃兰州 730046,中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, OIE/国家口蹄疫参考实验室, 甘肃兰州 730046,中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, OIE/国家口蹄疫参考实验室, 甘肃兰州 730046,中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, OIE/国家口蹄疫参考实验室, 甘肃兰州 730046,兰州大学基础医学院, 甘肃省新药临床前研究重点实验室, 甘肃兰州 730000,中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, OIE/国家口蹄疫参考实验室, 甘肃兰州 730046,中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, OIE/国家口蹄疫参考实验室, 甘肃兰州 730046,中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, OIE/国家口蹄疫参考实验室, 甘肃兰州 730046,中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, OIE/国家口蹄疫参考实验室, 甘肃兰州 730046,中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, OIE/国家口蹄疫参考实验室, 甘肃兰州 730046,中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, OIE/国家口蹄疫参考实验室, 甘肃兰州 730046,中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, OIE/国家口蹄疫参考实验室, 甘肃兰州 730046

基金项目：	甘肃省青年科技基金（1606RJYA256）；甘肃省自然科学基金（18JR3RA400）；中央级公益性科研院所基本科研业务费专项（1610312016018）

摘要：	【目的】为了研究O型口蹄疫病毒VP3G–H环中氨基酸突变对其生物学特性的影响。【方法】借助口蹄疫病毒反向遗传操作技术平台拯救出2株定点突变体rHN~(V3174Y)和rHN~(D3173N+V3174E+N3179C)。进行蚀斑形成试验、一步生长曲线的绘制、TCID_(50)和LD_(50)的测定、间接免疫荧光与激光共聚焦显微镜检测。【结果】结果显示,与骨架病毒rHN相比,虽然rHN~(V3174Y)和rHN~(D3173N+V3174E+N3179C)对BHK-21细胞的感染性及其蚀斑表型和复制动力学无显著性差异;但rHN~(V3174Y)和rHN~(D3173N+V3174E+N3179C)对乳鼠的致病力明显减弱,且均获得了小窝蛋白介导侵染CHO-K1细胞的能力。【结论】VP3上第3174位特征性氨基酸突变影响O型口蹄疫病毒感染宿主细胞的毒力及其内吞作用路径,这有助于我们认知VP3 G–H环在口蹄疫病毒粒子立体空间构象中潜在的作用。
关键词：	口蹄疫病毒 VP3 G-H环定点突变体生物学特性内吞作用
收稿时间：	2018/8/31 0:00:00
修稿时间：	2018/11/30 0:00:00
Effect of amino acid mutation(s) in the G-H loop of VP3 on biological characteristics of foot-and-mouth disease virus serotype O

Dongdong Chen,Xingwen Bai,Xiaohua Gong,Huifang Bao,Pinghua Li,Dong Li,Yuanfang Fu,Qifeng Bai,Hong Yuan,Pu Sun,Xueqing M,Yimei Cao,Yingli Chen,Zengjun Lu and Zaixin Liu.Effect of amino acid mutation(s) in the G-H loop of VP3 on biological characteristics of foot-and-mouth disease virus serotype O[J].Acta Microbiologica Sinica,2019,59(7):1285-1294.

Authors:	Dongdong Chen Xingwen Bai Xiaohua Gong Huifang Bao Pinghua Li Dong Li Yuanfang Fu Qifeng Bai Hong Yuan Pu Sun Xueqing M Yimei Cao Yingli Chen Zengjun Lu and Zaixin Liu

Institution:	State Key Laboratory of Veterinary Etiological Biology, OIE/China National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, China,State Key Laboratory of Veterinary Etiological Biology, OIE/China National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, China,State Key Laboratory of Veterinary Etiological Biology, OIE/China National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, China,State Key Laboratory of Veterinary Etiological Biology, OIE/China National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, China,State Key Laboratory of Veterinary Etiological Biology, OIE/China National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, China,State Key Laboratory of Veterinary Etiological Biology, OIE/China National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, China,State Key Laboratory of Veterinary Etiological Biology, OIE/China National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, China,Key Laboratory of Preclinical Study for New Drugs of Gansu Province, School of Basic Medical Sciences, Lanzhou University, Lanzhou 730000, Gansu Province, China,State Key Laboratory of Veterinary Etiological Biology, OIE/China National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, China,State Key Laboratory of Veterinary Etiological Biology, OIE/China National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, China,State Key Laboratory of Veterinary Etiological Biology, OIE/China National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, China,State Key Laboratory of Veterinary Etiological Biology, OIE/China National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, China,State Key Laboratory of Veterinary Etiological Biology, OIE/China National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, China,State Key Laboratory of Veterinary Etiological Biology, OIE/China National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, China and State Key Laboratory of Veterinary Etiological Biology, OIE/China National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu Province, China

Abstract:	Objective] To study the effect of amino acid mutation(s) in the G-H loop of VP3 of foot-and-mouth disease virus (FMDV) serotype O on its biological characteristics. Methods] Two site-directed mutants, rHN^V3174Y and rHN^{D3173N+V3174E+N3179C}, were generated by using reverse genetic technique platform of FMDV. The phenotypic properties of the resulting viruses were characterized by plaque forming assays, one-step growth curves, determination of TCID₅₀ and LD₅₀, indirect immunofluorescence assays and laser confocal microscopies. Results] There were no significant differences in the infectivity, plaque phenotype, and replication kinetics between rHN, rHN^V3174Y and rHN^{D3173N+V3174E+N3179C} for BHK-21 cells. Compared to the backbone virus rHN, however, the pathogenicity of rHN^V3174Y and rHN^{D3173N+V3174E+N3179C} was obvious decreased in suckling mice, and these two rescued viruses gained the ability to infect CHO-K1 cells via caveola-mediated endocytosis. Conclusion] Individual amino acid mutations at position 3174 in VP3 have an influence on the virulence and endocytic pathway of FMDV serotype O in the infection of host cells, which may help us understand the potentially important functional roles of the G-H loop of VP3 in the 3D spatial conformation of FMDV virion.

Keywords:	foot-and-mouth disease virus VP3 G-H loop site-directed mutant biological characteristics endocytosis
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