樱桃谷鸭源鹅出血性多瘤病毒VP3基因的克隆与序列分析

doi:10.16431/j.cnki.1671-7236.2017.04.006

《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (4): 980-985.doi: 10.16431/j.cnki.1671-7236.2017.04.006

樱桃谷鸭源鹅出血性多瘤病毒VP3基因的克隆与序列分析

万春和, 刘荣昌, 程龙飞, 傅光华, 傅秋玲, 施少华, 陈红梅, 黄瑜

福建省农业科学院畜牧兽医研究所, 福建省畜禽疫病防治工程技术研究中心, 福建省禽病防治重点实验室, 福州 350013

收稿日期:2016-08-01 出版日期:2017-04-20 发布日期:2017-05-03
通讯作者: 黄瑜 E-mail:huangyu_815@163.com
作者简介:万春和(1982-),男,湖北鄂州人,博士,副研究员,研究方向:水禽病毒分子生物学研究,E-mail:chunhewan@126.com
基金资助:
国家水禽产业技术体系（CARS-43）；国家自然科学基金（31602068）；福建省畜禽疫病防控技术重大研发平台（2014N2003）；福建省农业科学院畜牧兽医研究所基金（MYQJ2015（S）-3）；福建省属公益类项目（2015R1023-7）；福建省自然科学基金（2015J01114）

Cloning and Sequence Analysis of VP3 Gene of Cherry Valley Duck-origin Goose Hemorrhagic Polyomavirus

WAN Chun-he, LIU Rong-chang, CHENG Long-fei, FU Guang-hua, FU Qiu-ling, SHI Shao-hua, CHEN Hong-mei, HUANG Yu

Fujian Provincial Key Laboratory for Avian Diseases Control and Prevention, Fujian Animal Diseases Control Technology Development Center, Institute of Animal Husbandry and Veterinary Medicine of Fujian Academy of Agricultural Sciences, Fuzhou 350013, China

Received:2016-08-01 Online:2017-04-20 Published:2017-05-03

摘要/Abstract

摘要：

为明确福建地区是否存在鹅出血性多瘤病毒（goose hemorrhagic polyomavirus，GHPV）感染，本研究对2016年福建省临床送检测19份鸭组织样品进行GHPV检测，结果从1例樱桃谷鸭中检测到GHPV感染阳性（记为GHPV-FJ201601株）。随后根据GenBank中GHPV参考株序列特征，设计针对GHPV的VP3基因特异性引物，利用PCR技术扩增获得GHPV的VP3基因片段。结果显示，GHPV-FJ201601株的VP3基因全长为654 bp，编码217个氨基酸。对编码的VP3蛋白分析发现其理论等电点为9.37，带负电荷氨基酸为23个（Asp+Glu），带正电荷氨基酸为28个（Arg+Lys）；不稳定指数为38.43，是一个稳定蛋白；脂肪系数64.33，总平均疏水性指数为－0.744；该蛋白无典型的信号肽切割位点；其亚细胞定位类型为核定位；对其进行磷酸化分析发现，该蛋白存在14个氨基酸磷酸化位点。核苷酸同源性分析显示，不同来源GHPV代表株VP3基因相互之间核苷酸同源性均较高，均不低于99.7%。VP3基因遗传进化结果显示，GHPV-FJ201601株和鹅源GHPV分离株（匈牙利14234株与法国Toulouse Goose 2000株）处于同一遗传进化分支，与鸭源分离株（法国Toulouse Muscovy Duck 2008株、法国Toulouse Mule Duck 2008和中国106株）却处于不同的遗传进化分支，但所有GHPV分离株VP3基因遗传进化均较近。本研究首次证实福建地区樱桃谷鸭群中存在GHPV感染，为丰富不同地区与宿主的GHPV分子流行病学数据提供参考。

关键词: 鹅出血性多瘤病毒; 樱桃谷鸭; VP3基因; 克隆; 序列分析

Abstract:

In order to investigate goose hemorrhagic polyomavirus (GHPV) infection status in Fujian, China, 19 samples which were collected from different duck species were used to detecte GHPV, only one sample from Cheery Valley duck was tested positive (designated GHPV-FJ201601 strain). VP3 gene specific primers were designed by comparing the characterization of GHPV genomic sequences based on the genomic sequences download from GenBank. The length of the cloned VP3 gene was 654 bp, coding 217 amino acids. The theoretical pI of VP3 protein was 9.37, with 23 total number of negatively charged residues (Asp + Glu) and 28 total number of positively charged residues (Arg + Lys), respectively. The instability index of VP3 protein was 38.43, and it was a stable protein; The fat coefficient was 64.33; The total average hydrophobic index was -0.744. Also, the VP3 protein did not have signal peptides which was located in nucleus and having 14 phosphorylation sites. Nucleotides homology analysis of VP3 gene of GHPV isolates were higher with each other, which were no less than 99.7%. Phylogenetic analysis based on the VP3 gene showed that GHPV-FJ201601 strain shared the same genetic evolution cluster with goose-origin GHPV (14234 strain from Hungary and Toulouse Goose 2000 strain from France) rather than the duck-origin GHPV (Toulouse Muscovy Duck 2008 strain and Toulouse Mule Duck 2008 from France and 106 strain from China), all VP3 gene shared closer genetic distance with each other. In conclusion, this study was first evidence of Cheery Valley duck could infect GHPV in Fujian, which laid a good foundation for the enrichment the GHPV epidemiology data from different area and host.

Key words: goose hemorrhagic polyomavirus; Cherry Valley duck; VP3 gene; clone; sequence analysis

中图分类号:

S858.33

万春和, 刘荣昌, 程龙飞, 傅光华, 傅秋玲, 施少华, 陈红梅, 黄瑜. 樱桃谷鸭源鹅出血性多瘤病毒VP3基因的克隆与序列分析[J]. 《中国畜牧兽医》, 2017, 44(4): 980-985.

WAN Chun-he, LIU Rong-chang, CHENG Long-fei, FU Guang-hua, FU Qiu-ling, SHI Shao-hua, CHEN Hong-mei, HUANG Yu. Cloning and Sequence Analysis of VP3 Gene of Cherry Valley Duck-origin Goose Hemorrhagic Polyomavirus[J]. , 2017, 44(4): 980-985.

参考文献

[1] Lacroux C,Andreoletti O,Payre B,et al. Pathology of spontaneous and experimental infections by goose haemorrhagic polyomavirus[J]. Avian Pathol,2004,33(3):351-358.
[2] Dobos-Kovács M,Horváth E,Farsang A,et al. Haemorrhagic nephritis and enteritis of geese:Pathomorphological investigations and proposed pathogenesis[J]. Acta Vet Hung,2005,53(2):213-223.
[3] Bernath S,Szalai F. Investigations for clearing the etiology of disease appeared among gostlings in 1969[J]. Magyar Alla Lap,1970,25:531-536.
[4] Guerin J,Gelfi J,Dubois L,et al. A novel polyomavirus (goose hemorrhagic polyomavirus) is the agent of hemorrhagic nephritis enteritis of geese[J]. J Virol,2000,74(10):4523-4529.
[5] Johne R,Müller H. The genome of goose hemorrhagic polyomavirus,a new member of the proposed subgenus Avipolyomavirus[J]. Virology,2003,308(2):291-302.
[6] Gawe? A,Woz'niakowski G,Samorek-Salamonowicz E,et al. Hemorrhagic nephritis and enteritis in a goose flock in Poland——Disease course analysis and characterization of etiologic agent[J]. Avian Dis,2014,58(4):518-522.
[7] Bernáth S,Farsang A,Kovács A,et al. Pathology of goose haemorrhagic polyomavirus infection in goose embryos[J]. Avian Pathol,2006,35(1):49-52.
[8] Pingret J,Boucraut-Baralon C,Guérin J L. Goose haemorrhagic polyomavirus infection in ducks[J]. Vet Rec,2008,162(5):164.
[9] Corrand L,Gelfi J,Albaric O,et al. Pathological and epidemiological significance of goose haemorrhagic polyomavirus infection in ducks[J]. Avian Pathol,2011,40(4):355-360.
[10] 姜甜甜,张大丙.北京鸭源鹅出血性多瘤病毒的分子检测[J].中国兽医杂志,2012,48(6):3-6.
[11] Rychlik W. OLIGO 7 primer analysis software[J]. Methods in Molecular Biology,2007,402:35-60.
[12] 聂鑫,赵天靖,曹瑞勇,等.羊种布鲁氏菌LpxB基因的克隆、原核表达及其蛋白的生物信息学分析[J]. 中国畜牧兽医,2016,43(7):1681-1687.
[13] 杨玉静,聂瑞强,谢建山,等.小鼠Oct-1基因CDS区克隆与生物信息学分析[J].中国畜牧兽医,2016,43(8):1960-1966.
[14] 赵孟孟,宋中宝,冯松林,等.猪繁殖与呼吸综合征病毒FS株Nsp9基因的克隆与序列分析[J]. 中国畜牧兽医,2016,43(9):2249-2256.
[15] Liu H,Wang H,Tian X,et al. Complete genome sequence of goose parvovirus Y strain isolated from Muscovy ducks in China[J]. Virus Genes,2014,48(1):199-202.
[16] 黄瑜,万春和,傅秋玲,等.新型番鸭细小病毒的发现及其感染的临床表现[J].福建农业学报,2015,30(5):442-445.
[17] Chen H,Dou Y,Tang Y,et al. Genomic characterization of a duck-origin gpv-related parvovirus from Cherry Valley ducklings in China[J]. PLoS One,2015,10(10):e0140284.
[18] Chen S,Wang S,Cheng X,et al. Isolation and characterization of a distinct duck-origin goose parvovirus causing an outbreak of duckling short beak and dwarfism syndrome in China[J]. Arch Virol,2016,161(9):2407-2416.
[19] 万春和,陈红梅,傅秋玲,等.番鸭细小病毒浙江分离株VP基因的克隆与序列分析[J]. 中国畜牧兽医,2015,42(10):2600-2605.
[20] Liu M,Meng F,Li X,et al. Goose haemorrhagic hepatitis caused by a new subtype duck hepatitis type 1 virus[J]. Vet Microbiol,2011,152(3-4):280-283.
[21] Shehata A,Gerry D,Heenemann K,et al. Goose parvovirus and circovirus coinfections in ornamental ducks[J]. Avian Dis,2016,60(2):516-522.

樱桃谷鸭源鹅出血性多瘤病毒VP3基因的克隆与序列分析

Cloning and Sequence Analysis of VP3 Gene of Cherry Valley Duck-origin Goose Hemorrhagic Polyomavirus

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