2株猪繁殖与呼吸综合征病毒海南株的ORF5基因分析

doi:10.6048/j.issn.1001-4330.2023.12.021

新疆农业科学 ›› 2023, Vol. 60 ›› Issue (12): 3049-3056.DOI: 10.6048/j.issn.1001-4330.2023.12.021

• 畜牧兽医·微生物·农业装备工程与机械化 • 上一篇下一篇

2株猪繁殖与呼吸综合征病毒海南株的ORF5基因分析

马佳镁¹^,²(), 钟植文³, 范悦轩¹^,², 黄春媛², 郑佳馨², 曹芳芳², 王金泉¹, 刘光亮¹^,⁴(), 曹宗喜¹^,²()

1.新疆农业大学动物医学学院,乌鲁木齐 830052
2.海南省农业科学院畜牧兽医研究所/海南省热带动物繁育与疫病研究重点实验室/国家动物疫病数据中心海口观测实验点,海口 571100
3.广东永顺生物制药股份有限公司,广州 511536
4.海南省农业科学院畜牧兽医研究所海南省院士团队创新中心,海口 571100

收稿日期:2023-04-08 出版日期:2023-12-20 发布日期:2024-01-03
通信作者: 曹宗喜(1982-),男,河南新乡人,研究员,博士,硕士生导师,研究方向为家畜传染病学,(E-mail)caozongxi@163.com;刘光亮(1975- ),男,四川资中人,研究员,博士,博士生导师,研究方向为动物疫苗研制及分子免疫学,(E-mail)liuguangliang01@caas.cn
作者简介:马佳镁(1997-),女,甘肃庆阳人,硕士研究生,研究方向为家畜传染病学,(E-mail)2452628323@qq.com
基金资助:
海南省省属科研院所技术创新专项(jscx202001);国家自然科学基金项目(32060796);家畜疫病病原生物学国家重点实验室开放基金(SKLVEB2020KFKT021);海南省农业科学院院本级科研项目(HAAS2022PT0205);国家重点研发计划(2023YFC3404302)

ORF5 gene analysis of two porcine reproductive and respiratory syndrome virus strains from hainan province

MA Jiamei¹^,²(), ZHONG Zhiwen³, FAN Yuexuan¹^,², HUANG Chunyuan², ZHENG Jiaxin², CAO Fangfang², WANG Jinquan¹, LIU Guangliang¹^,⁴(), CAO Zongxi¹^,²()

1. College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, China
2. Institute of Animal Husbandry and Veterinary Medicine,Hainan Academy of Agricultural Sciences/Hainan Provincial Key Laboratory of Tropical Animal Breeding and Epidemic Research in Hanan Province/ Haikou Observation Experimental Station of National Data Center of Animal Health, Haikou 571100, China
3. Guangdong Winsun Bio-Pharmaceutical Co.,Ltd., Guangzhou 511536, China
4. Hainan Provincial Academician Innovation Center, Institute of Animal Husbandry and Veterinary Medicine, Hainan Academy of Agricultural Sciences, Haikou 571100, China

Received:2023-04-08 Online:2023-12-20 Published:2024-01-03
Correspondence author: CAO Zongxi(1982-),male,XinXiang Henan,Master Tutor,fellows,Ph.D., research in livestock epidemiology,(E-mail)caozongxi@163.com;LIU Guangliang(1982-),male,Doctoral supervisor,fellows,Ph.D.,research direction of animal vaccine development and molecular immunology,(E-mail)liuguangliang01@caas.cn
Supported by:
Technological Innovation Project of Hainan Provincial Scientific Research Institutes(jscx202001);National Natural Science Foundation of China(32060796);Open Fund of the State Key Laboratory of Pathogenic Biology of Livestock Diseases(SKLVEB2020KFKT021);Hainan Provincial Academy of Agricultural Sciences Research Project(HAAS2022PT0205);National key Research and Development Program of China(2023YFC3404302)

摘要/Abstract

摘要：

【目的】鉴定我国海南省新发现的2株猪繁殖与呼吸综合征病毒株HNHK1-2021和HNLG1-2021的进化关系。【方法】采用PCR方法对ORF5基因进行扩增和序列测定,利用MegAlign软件进行多重序列比对,利用MEGA X软件邻接法构建进化树。【结果】PRRSV HNHK1-2021和HNLG1-2021毒株核苷酸相似性为82.9%,推导氨基酸相似性为82.6%。HNHK1-2021毒株和NADC30类 PRRSV、QYYZ类 PRRSV、VR-2332类PRRSV以及JXA1类PRRSV序列比对,80.8% ~ 82.8%、90.2% ~ 92.0%、82.1%~83.4%、81.9%~84.4%,推导氨基酸相似性分别为82.1% ~ 84.6%、91.5% ~ 94.5%、80.1% ~ 83.1%、82.6% ~ 84.6%;HNLG1-2021毒株和NADC30类PRRSV、QYYZ类PRRSV、VR2332类PRRSV以及JXA1类PRRSV序列比对,核苷酸相似性分别为83.7% ~ 84.6%、81.8% ~ 83.6%、97.7% ~ 99.5%、85.6% ~ 90.4%,推导氨基酸相似性为83.6% ~ 85.1%、81.1% ~ 82.6%、96.0% ~ 98.5%,86.1% ~ 90.5%。【结论】2株病毒株均属于北美型PRRSV,HNHK1-2021毒株属于QYYZ类PRRSV(谱系3)。HNLG1-2021毒株属于VR-2332类PRRSV(谱系5)。

关键词: 猪繁殖与呼吸综合征病毒; ORF5基因; 多重序列比对; 同源性

Abstract:

【Objective】This project aims to identify the evolutionary relationship of two newly discovered porcine reproductive and respiratory syndrome virus strains HNHK1-2021 and HNLG1-2021 in Hainan,China.【Methods】The ORF5 gene was amplified and sequenced by PCR method. Multiple sequence alignment was performed using MegAlign software, and phylogenetic tree was constructed using MEGA X software neighbor joining method.【Results】The nucleotide similarity of PRRSV HNHK1-2021 and HNLG1-2021 strains was 82.9%, and the deduced amino acid similarity was 82.6%. HNHK1-2021 strain and NADC30-like PRRSV, QYYZ-like PRRSV, VR2332-like PRRSV and JXA1-like PRRSV sequence alignment: 80.8%-82.8%, 90.2%-92.0%, 82.1%-83.4%, 81.9%-84.4%,deducing the amino acid similarity was 82.1%-84.6%, 91.5%-94.5%, 80.1%-83.1%, 82.6%-84.6%; HNLG1-2021 strain and NADC30 PRRSV, QYYZ PRRSV, VR2332 PRRSV and JXA1 PRRSV sequence alignment, the nucleotide similarity of which was 83.7%-84.6%、81.8%-83.6%, 97.7%-99.5%, 85.6%-90.4%, the deduced amino acid similarity was 83.6%-85.1%, 81.1%-82.6%, 96%-98.5%, 86.1%-90.5%.【Conclusion】The two virus strains belong to the North American type PRRSV, and the HNHK1-2021 strain belongs to the QYYZ class PRRSV (lineage 3). The HNLG1-2021 strain belongs to the VR-2332 class of PRRSVs (lineage 5).

Key words: porcine reproductive and respiratory syndrome virus; ORF5 gene; multiple sequence alignment; homology

中图分类号:

S855.2

马佳镁, 钟植文, 范悦轩, 黄春媛, 郑佳馨, 曹芳芳, 王金泉, 刘光亮, 曹宗喜. 2株猪繁殖与呼吸综合征病毒海南株的ORF5基因分析[J]. 新疆农业科学, 2023, 60(12): 3049-3056.

MA Jiamei, ZHONG Zhiwen, FAN Yuexuan, HUANG Chunyuan, ZHENG Jiaxin, CAO Fangfang, WANG Jinquan, LIU Guangliang, CAO Zongxi. ORF5 gene analysis of two porcine reproductive and respiratory syndrome virus strains from hainan province[J]. Xinjiang Agricultural Sciences, 2023, 60(12): 3049-3056.

图/表 6

表1 引物序列信息

Tab.1 Primer sequence information

基因 Gene	引物名称 Name	引物序列 Primer sequences (5’→3’)	扩增片段大小 Fragment length (bp)
ORF5	ORF5-F	5'GTTTTAGCCTGTCTTTTTGCC3'	731
ORF5	ORF5-R	5' TATATCATCACTGGCGTGTAGG3'	731

表2 PRRSV代表毒株信息

Tab.2 Information of representative PRRSV strains

编号 Number	毒株名称 Strains	年份 Year	登录号 GenBank accession NO.
1	BJ-4	2000	AF331831
2	CH-1a	2000	AY032626
3	PA8	2002	AF176348
4	HN1	2003	AY457635
5	LU534	2004	AY615794
6	JAX1	2006	EF112445
7	HUN4	2007	EF635006
8	VR2332	2007	U87392
9	HaiNan-1	2007	EF398051
10	HaiNan-2	2007	EF398056
11	XH-GD	2008	EU624117
12	GS2003	2009	EU880442
13	NADC30	2011	JN654459
14	QYYZ	2011	JQ308798
15	YJ1-10	2012	KC282627
16	HENAN-XINX	2013	KF611905
17	HNHK2	2014	KY488475
18	HNHK1	2014	KY488474
19	FJZ03	2015	KP860909
20	FJYR	2015	KT804696
21	HNjz15	2016	KT945017
22	JX143	2016	EU708726
23	GDZJ1	2017	KY508635
24	GXBB130624	2017	MF807236
25	JS201411	2018	MF095210
26	GZgy17	2018	MK144542
27	10186-614	2020	LC569874

图1 ORF5基因琼脂糖凝胶电泳结果注:M:DL-2 000 DNA Marker;1:HNHK1-2021毒株ORF5;2.HNLG1-2021 毒株ORF5;3.阴性对照

Fig.1 ORF5 gene agarose gel electrophoresis results Note: M: DL-2 000 DNA Marker; 1: HNHK1-2021 strain ORF5; 2.HNLG1-2021 strain ORF5; 3.Negative control

表3 ORF5基因核苷酸(右上方)和氨基酸序列(左下方)的同源性

Tab.3 Homology analysis results of ORF5 gene nucleotide (upper right) and amino acid sequence (lower left)

图2 PRRSV ORF5氨基酸序列比对

Fig.2 PRRSV ORF5 amino acid sequence alignment

图3 ORF5基因遗传进化树注: 为本次分离得到的毒株, 为海南不同时期分离得到的毒株

Fig.3 ORF5 gene genetic tree Note:The strain obtained from this isolation,Viras s trains is olated from different Periods in Hainan Province

参考文献 21

[1]	Pileri E, Mateu E. Review on the transmission porcine reproductive and respiratory syndrome virus between pigs and farms and impact on vaccination[J]. Vet Res. 2016, 47(1):108. PMID
[2]	杨汉春, 周磊. 猪繁殖与呼吸综合征病毒的遗传变异与演化[J]. 生命科学, 2016, 28(3):325-336.
	YANG Hanchun, ZHOU Lei. Genetic variation and evolution of porcine reproductive and respiratory syndrome virus[J]. Chinese Bulletin of Life Sciences, 2016, 28(3):325-336.
[3]	Zhao J, Zhu L, Deng H, et al. Genetic characterization of a novel porcine reproductive and respiratory syndrome virus type I strain from southwest China[J]. Archives of Virology, 2021, 166(6):1769-1773. DOI PMID
[4]	欧阳艳, 周艳荣, 方六荣, 等. 我国生猪集中养殖区PRRSV流行及遗传特征分析[J]. 动物医学进展, 2022, 345(3):33-37.
	OUYANG Yan, ZHOU Yanrong, FANG Liurong, et al. Analysison the Prevalence and Genetic Characteristics of PRRSV in Pig Concentrated Breeding Areas of China[J]. Progress in Veterinary Medicine, 2022, 345(3):33-37.
[5]	Cavanagh D. Nidovirales: a new order comprising Coronaviridae and Arteriviridae[J]. Archives of Virology, 1996, 142(3):629-633.
[6]	Snijder E J, Kikkert M, Fang Y. Arterivirus molecular biology and pathogenesis[J]. Journal of General Virology, 2013, 94(10):2141-2163.. DOI URL
[7]	Cao Z X, Jiao P R, Huang Y M, et al. Genetic diversity analysis of the ORF5 gene in porcine reproductive and respiratory syndrome virus samples from South China[J]. Acta Veterinaria Hungarica, 2012, 60(1):157-164. DOI URL
[8]	贺冬梅, 谭树义, 陈朝喜, 等. PRRSV HuN-2011株ORF5和ORF7基因的序列分析[J]. 黑龙江畜牧兽医, 2017,(7):188-191.
	HE Dongmei, TAN Shuyi, CHEN Zhaoxi, et al. Sequence Analysis of ORF5 and ORF7 Gene of PRRSV HuN-2011 Strain[J]. Heilongjiang Animal Husbandry and Veterinary Medicine, 2017,(7):188-191.
[9]	Shi M, Tommy T L, Chung-Chau H, et al. Molecular epidemiology of PRRSV:A phylogenetic perspective[J]. Virus Research, 2010, 154(1-2):7-17. DOI URL
[10]	Liu J, Xu Y, Lin Z. et al. Epidemiology investigation of PRRSV discharged by faecal and genetic variation of ORF5[J]. Transboundary and Emerging Diseases, 2021, 68(4):2334-2344. DOI PMID
[11]	Ma H, Li X, Li J, et al. Immunization with a recombinant fusion of porcine reproductive and respiratory syndrome virus modified GP5 and ferritin elicits enhanced protective immunity in pigs[J]. Virology, 2021, 552:112-120. DOI PMID
[12]	陆桂丽, 汪萍, 成进, 等. 猪繁殖与呼吸综合征新疆株ORF2-7基因的克隆及序列分析[J]. 新疆农业科学, 2014, 51(3):546-551.
	LU Guili, WANG Ping, CHENG Jin, et al. Clone Andand Sequence Analysis of ORF2-7 Genes of Porcine Reproductive and Respiratory Syndrome Virus of Xinjiang Strains[J]. Xinjiang Agricultural Sciences, 2014, 51(3):546-551.
[13]	Kick A R, Amaral A F, Diego A, et al. The local and systemic humoral immune response against homologous and heterologous strains of the type 2 Porcine Reproductive and Respiratory Syndrome Virus[J]. Frontiers in Immunology, 2021.
[14]	郭宝清, 陈章水, 刘文兴, 等. 从疑似PRRS流产胎儿分离PRRSV的研究[J]. 中国预防兽医学报, 1996,(2):1-4.
	GUO Baoqing, CHEN Zhangshui, LIU Wenxing, et al. Study on Isolation of PRRSV from Aborted Fetuses Suspected of PRRS[J]. Chinese Journal of Preventive Veterinary Medicine, 1996,(2):1-4.
[15]	Cao Z X, Pan Q H, Lin Z M, et al. Isolation and sequence analysis of NSP2, ORF3 and ORF5 of PRRSV XH-GD strain from Guangdong province[J]. Agricultural Biotechnology, 2014, 3(1):4-8.
[16]	Ruedas-Torres I, Rodriguze-Gomez I M, Sanchez-Carvajal J M, et al. The jigsaw of PRRSV virulence[J]. Veterinary Microbiology, 2021, 260.
[17]	Zhou L, Kang R M, Zhang Y, et al. Emergence of two novel recombinant porcine reproductive and respiratory syndrome viruses 2 (lineage 3) in Southwestern China[J]. Veterinary Microbiology, 2019, 232:30-41. DOI PMID
[18]	Den Boon JA, Faaberg KS, Meulenberg JJ, et al. Processing and evolution of the N-terminal region of the arterivirus replicase ORF1a protein: identification of two papainlike cysteine proteases[J]. Journal of Virology, 1995, 69(7):4500-4505. PMID
[19]	Li B, Xiao S, Wang Y, et al. Immunogenicity of the highly pathogenic porcine reproductive and respiratory syndrome virus GP5 protein encoded by a synthetic ORF5 gene[J]. Vaccine, 2009, 27(18):1957-1963. DOI URL
[20]	Barfoed A M, Blixenkrone-Moller M, Jensen M H, et al. DNA vaccination of pigs with open reading frame 1-7 of PRRS virus[J]. Vaccine, 2004, 22(27-28):3628-3641. PMID
[21]	Yu L, Zhao P, Dong J, et al. Genetic characterization of 11 porcine reproductive and respiratory syndrome virus isolates in South China from 2014 to 2015[J]. Journal of Virology, 2017, 14(1):139. DOI URL

2株猪繁殖与呼吸综合征病毒海南株的ORF5基因分析

ORF5 gene analysis of two porcine reproductive and respiratory syndrome virus strains from hainan province

在线阅读

PDF下载

可视化

摘要/Abstract

引用本文

使用本文

图/表 6

参考文献 21

相关文章 6

编辑推荐

Metrics

[1]	吴鹏;张勋;梁田;陈新凯;肖媛媛;盛金良. 北疆地区猪繁殖与呼吸综合征病毒ORF7基因克隆与遗传演化分析[J]. , 2016, 53(11): 2149-2156.
[2]	陆桂丽;汪萍;成进;夏俊;沙依兰·卡依扎;吐尔洪·努尔;王文. 猪繁殖与呼吸综合征新疆株ORF2-7基因的克隆及序列分析[J]. , 2014, 51(3): 546-551.
[3]	高静涛;牛建新;王雪莲;刘娜;叶春秀;张飞;朱天丁. 枣疯病植原体的分子检测与同源性鉴定[J]. , 2011, 48(4): 662-667.
[4]	李谨;杨公社;陈创夫. 新疆军垦型细毛羊MHC基因临近区段3个遗传标记的研究[J]. , 2008, 45(6): 1182-1186.
[5]	苟荣;姚新奎;李红;谭晓海;努尔巴哈提;马江飞;刘珩;玉山江;叶再华;陈阳;潘晓东. 酸马奶中酵母菌5.8 S rDNA及ITS的基因序列分析[J]. , 2008, 45(5): 868-874.
[6]	李永丹;史雪岩;田兆丰;阿不都外力·依玛木;高希武. 新疆红胫戟纹蝗痘病毒包涵体蛋白基因序列分析[J]. , 2006, 43(1): 38-42.