鹦鹉幼雏病病毒福建株VP1基因的克隆及序列分析

doi:10.16431/j.cnki.1671-7236.2018.06.030

《中国畜牧兽医》 ›› 2018, Vol. 45 ›› Issue (6): 1661-1667.doi: 10.16431/j.cnki.1671-7236.2018.06.030

鹦鹉幼雏病病毒福建株VP1基因的克隆及序列分析

万春和, 程龙飞, 傅光华, 施少华, 陈红梅, 傅秋玲, 刘荣昌, 林建生, 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:2017-12-06 出版日期:2018-06-20 发布日期:2018-06-15
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:水禽病原分子生物学,E-mail:chunhewan@126.com
基金资助:
福建省农业科学院科技创新团队（STIT2017-3-10、STIT2017-1-5）；国家水禽产业体系（CARS-42）；国家自然科学基金（31602068）；福建省属公益类项目（2018R1023-5）；福建省农业科学院青年科技英才项目（YC2015-12）

Cloning and Sequence Analysis of VP1 Gene of Budgerigar Fledgling Disease Polyomavirus from Fujian

WAN Chunhe, CHENG Longfei, FU Guanghua, SHI Shaohua, CHEN Hongmei, FU Qiuling, LIU Rongchang, LIN Jiansheng, 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:2017-12-06 Online:2018-06-20 Published:2018-06-15

摘要/Abstract

摘要：

为明确虎皮鹦鹉源鹦鹉幼雏病病毒（budgerigar fledgling disease polyomavirus，BFPyV）福建株（命名为FJ-2016株）结构蛋白VP1基因特征，本研究针对BFPyV VP1基因特点设计特异性引物，利用PCR方法扩增获得FJ-2016株VP1基因全长序列，目的片段经胶回收后进行克隆测序。结果显示，BFPyV FJ-2016株VP1基因全长为1 032 bp，编码343个氨基酸。所编码VP1蛋白的理论等电点为5.77，不稳定指数为40.91，是不稳定蛋白；脂肪系数为74.72；总平均疏水性指数为-0.366。将获得的VP1基因序列提交至GenBank，登录号为：MG148345。核苷酸同源性分析表明，不同时间、地区和品种BFPyV的VP1基因十分保守，相互之间的核苷酸同源性均在99.1%以上。遗传进化分析发现，不同时间和地域BFPyV相互之间亲缘关系较近，但可细分为两个大的遗传进化分支（Clade 1和Clade 2）。从宿主品种来看，虎皮鹦鹉源BFPyV各分离株的遗传进化与分离时间及地域无明显关系，虎皮鹦鹉源BFPyV分离株在Clade 1和Clade 2分支均有分布。本研究首次证实福建地区虎皮鹦鹉中存在BFPyV感染，相关研究结果为丰富不同地区BFPyV分子流行病学数据提供参考。

关键词: 鹦鹉幼雏病病毒; 虎皮鹦鹉; VP1基因; 序列分析

Abstract:

In order to characterize the VP1 gene of budgerigar fledgling disease polyomavirus (BFPyV) from Melopsittacus undulates (designated as FJ-2016 strain),specific primers were designed according to the VP1 gene of BFPyV retrieved from GenBank.The full-length sequence of VP1 gene of FJ-2016 strain was amplified by PCR.The target fragment was recovered,cloned and sequenced.The results demonstrated that the length of VP1 gene was 1 032 bp,coding 343 amino acids.The VP1 protein theoretical pI,instability index,aliphatic index and grand average of hydropathicity was 5.77,40.91(means unstable protein),74.72 and -0.366,respectively.The sequences were submitted to the GenBank,the accession No. was MG148345.Nucleotides homology of the VP1 gene of BFPyV isolates were above 99.1% with each other.The VP1 gene nucleotides comparison indicated that the VP1 gene was very conserved among different time,regions and species.Phylogenetic tree based on the VP1 gene showed that BFPyV isolates with different time and regions shared closer relationship with each other,which also could be divided into two genetic clades (Clade 1 and Clade 2).The Melopsittacus undulates origin BFPyV could be found in the two genetic clades,these data indicated that Melopsittacus undulates origin BFPyV shared no significant phylogenetic evolution relationship with the time and regions.In conclusion,this study provided evidence of budgerigar fledgling disease polyomavirus circulating in Fujian,which enriched the BFPyV molecular epidemiology data in China.

Key words: budgerigar fledgling disease polyomavirus; Melopsittacus undulatus; VP1 gene; sequence analysis

中图分类号:

S858.93

万春和, 程龙飞, 傅光华, 施少华, 陈红梅, 傅秋玲, 刘荣昌, 林建生, 黄瑜. 鹦鹉幼雏病病毒福建株VP1基因的克隆及序列分析[J]. 《中国畜牧兽医》, 2018, 45(6): 1661-1667.

WAN Chunhe, CHENG Longfei, FU Guanghua, SHI Shaohua, CHEN Hongmei, FU Qiuling, LIU Rongchang, LIN Jiansheng, HUANG Yu. Cloning and Sequence Analysis of VP1 Gene of Budgerigar Fledgling Disease Polyomavirus from Fujian[J]. , 2018, 45(6): 1661-1667.

参考文献

[1] 寇铮.鹦鹉幼雏病病毒全序列分析及VP1基因克隆与表达[D].武汉:华中农业大学,2004. KOU Z.Complete sequence analysis of budgerigar fledgling disease virus and cloning and expression of VP1 gene[D].Wuhan:Huazhong Agricultural University,2004.(in Chinese)
[2] 吴延功,张子春,王玉东,等.中国发现澳洲长尾小鹦鹉幼雏病[J].中国兽医学报,1996,16(3):218-221. WU Y G,ZHANG Z C,WANG Y D,et al.Budgerigar fledgling disease occurred in China[J]. Chinese Journal of Veterinary Science,1996,16(3):218-221.(in Chinese)
[3] JOHNE R,MVLLER H.Avian polymavirus in wild birds:Genome analysis of isolates from Falconiformes and Psittaciformes[J].Archives of Virology,1998,143(8):1501-1512.
[4] ROTT O,KRÖGER M,MVLLER H,et al.The genome of budgerigar fledgling disease virus,an avian polyomavirus[J].Virology,1988,165(1):74-86.
[5] STOLL R,LUO D,KOUWENHOVEN B,et al.Molecular and biological characteristics of avian polyomaviruses:Isolates from different species of birds indicate that avian polyomaviruses form a distinct subgenus within the polyomavirus genus[J].Journal of General Virology,1993,74(2):229-237.
[6] STOLL R,HOBOM G,MVLLER H.Host restriction in the productive cycle of avian polyomavirus budgerigar fledgling disease virus type 3 depends on a single amino acid change in the common region of structural proteins VP2/VP3[J].Journal of General Virology,1994,75(9):2261-2269.
[7] LUO D,MVLLER H,TANG X,et al.Early and late pre-mRNA processing of budgerigar fledgling disease virus 1:Identification of viral RNA 5' and 3' ends and internal splice junctions[J].Journal of General Virology,1995,76(1):161-166.
[8] LI J,LIU Q,MVLLER H,et al.Avian polyomavirus expression patterns of bicistronic late mRNAs[J].Virology,2009,388(1):42-48.
[9] MOENS U,KRUMBHOLZ A,EHLERS B,et al.Biology,evolution,and medical importance of polyomaviruses:An update[J].Infection Genetics and Evolution,2017,54:18-38.
[10] MOENS U,CALVIGNAC-SPENCER S,LAUBER C,et al.ICTV virus taxonomy profile:Polyomaviridae[J].Journal of General Virology,2017,98(6):1159-1160.
[11] BOZEMAN L H,DAVIS R B,GAUDRY D,et al.Characterization of a papovavirus isolated from fledgling budgerigars[J].Avian Disases, 1981,25(4):972-980.
[12] LEHN H,MVLLER H.Cloning and characterization of budgerigar fledgling disease virus,an avian polyomavirus[J].Virology,1986,151(2):362-370.
[13] KATOH H,OGAWA H,OHYA K,et al.A review of DNA viral infections in psittacine birds[J].The Journal of Veterinary Medicine Science,2010,72(9):1099-1106.
[14] 夏苇,冯峰,王旭,等.我国一株鹦鹉新病毒的理化性质及鉴定[J].华中师范大学学报(自然科学版),2000,34(2):216-219. XIA W,FENG F,WANG X,et al.Phychemical characteristic and identification of a new psittacine virus in China[J]. Journal of Central China Normal University (Nature Science Edition),2000,34(2):216-219.(in Chinese)
[15] 庄青叶,蒋文明,侯广宇,等.山东一株鹦鹉幼雏病病毒全序列测定与分析[J].现代生物医学进展,2009,9(10):1885-1887. ZHUANG Q Y,JIANG W M,HOU G Y,et al.Sequencing and analysis of complete genome of budgerigar fledgling disease virus isolated from Shandong province[J]. Progress in Modern Biomedicine,2009,9(10):1885-1887.(in Chinese)
[16] JOHNE R,MVLLER H.Polyomaviruses of birds:Etiologic agents of inflammatory diseases in a tumor virus family[J].Journal of Virology,2007,81(21):11554-11559.
[17] AMBALATHINGAL G R,FRANCIS R S,SMYTH M J,et al.BK polyomavirus:Clinical aspects,immune regulation,and emerging therapies[J].Clinical Microbiology Reviews, 2017,30(2):503-528.
[18] KOU Z,ZHANG Z,CHEN S,et al.Molecular characterizations of avian polyomavirus isolated from budgerigar in China[J].Avian Diseases,2008,52(3):451-454.
[19] DAYARAM A,PIASECKI T,CHRZA?TEK K,et al.Avian polyomavirus genome sequences recovered from parrots in captive breeding facilities in Poland[J].Genome Announcements,2015,3(5):e00986-15.
[20] KATOH H,OHYA K,UNE Y,et al.Molecular characterization of avian polyomavirus isolated from psittacine birds based on the whole genome sequence analysis[J].Veterinary Microbiology,2009,138(1-2):69-77.
[21] ZHUANG Q,CHEN J,MUSHTAQ M H,et al.Prevalence and genetic characterization of avian polyomavirus and psittacine beak and feather disease virus isolated from budgerigars in Mainland China[J].Archives of Virology,2012,157(1):53-61.
[22] PERPIÑÁN D,GARNER M M,WELLEHAN J F,et al.Mixed infection with reovirus and Chlamydo-phila in a flock of budgerigars (Melopsittacus undulatus)[J].Journal of Avian Medicine and Surgery, 2010,24(4):316-321.
[23] IMADA T,YAMAGUCHI S,KAWAMURA H,et al.Isolation of an influenza A virus from the budgerigar,Melopsittacus undulatus[J].National Institute of Animal Health Quarterly (Tokyo),1980,20(1):30-31.
[24] WAJID A,BASHARAT A,KHAN T A,et al.Complete genome sequence of a velogenic Newcastle disease virus strain isolated from a clinically healthy exotic parakeet (Melopsittacus undulatus) in Pakistan[J].Genome Announcements,2017,5(6):e01581-16.
[25] CUI J,HOLMES E C.Endogenous hepadnaviruses in the genome of the budgerigar (Melopsittacus undulatus) and the evolution of avian hepadnaviruses[J].Journal of Virology,2012,86(14):7688-7691.
[26] 万春和,刘荣昌,程龙飞,等.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 C H,LIU R C,CHENG L F,et al.Cloning and sequence analysis of VP3 gene of Cherry Valley duck-origin goose hemorrhagic polyomavirus[J].China Animal Husbandry & Veterinary Medicine, 2017,44(4):980-985.(in Chinese)
[27] BARON H R,HOWE L,VARSANI A,et al.Disease screening of three breeding populations of adult exhibition budgerigars (Melopsittacus undulatus) in New Zealand reveals a high prevalence of a novel polyomavirus and avian malaria infection[J].Avian Diseases,2014,58(1):111-117.

鹦鹉幼雏病病毒福建株VP1基因的克隆及序列分析

Cloning and Sequence Analysis of VP1 Gene of Budgerigar Fledgling Disease Polyomavirus from Fujian

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