基于N基因的小反刍兽疫病毒贵州流行株分子特征与分群研究

doi:10.16431/j.cnki.1671-7236.2018.08.003

Abstract

Abstract:

In order to investigate the N gene molecular characteristics and clustering of the peste des petits ruminants virus (PPRV) of Guizhou province,the primers of PPRV N gene were designed and the target gene was amplified from clinical samples and then was cloned to pMD19-T vector.The positive recombinant plasmids were sequenced and nucleotide homologoy, amino acids homology,mutation sites and phylogenetic tree analysis of N gene of Guizhou PPRV strains and reference strains were performed by DANStar software.The results showed that the length of amplified N gene was 1 578 bp and the homology of nucleotide and amino acid sequences among Guizhou PPRV strains were 99.6% to 100.0% and 99.2% to 100.0%,respectively.The homology of nucleotide (97.7% to 99.9%) and amino acids (98.3% to 100.0%) compared with domestic reference strains were higher than those of other countries (88.5% to 97.7% and 92.2% to 98.5%).Compared with the vaccine strain Nigeria 75-1,there were 26 multiple sites in N gene of Guizhou PPRV strains while there was no amino acid loss or increase.The results of phylogenetic analysis showed that Guizhou PPRV strains were in the same evolutionary branch with the domestic reference strains,while the foreign reference strains in a different evolutionary branch.Guizhou PPRV strains belonged to Ⅳ gene group and vaccine strain Nigeria 75-1 belonged to Ⅰ gene group.

Key words: peste des petits ruminants virus (PPRV); N gene; sequence analysis; gene grouping

CLC Number:

S852.65

WANG Jun, YANG Yuan, ZHANG Yundan, WEN Ming, ZHOU Bijun, CHENG Zhentao, YUN Jun, LI Tao. Molecular Characteristics and Cluster of PPRV Guizhou Strains Based on N Gene[J]. , 2018, 45(8): 2057-2066.

References

[1] ABUBAKAR M,JAMAL S M,ARSHED M J,et al.Peste des petits ruminants virus (PPRV) infection;Its association with species,seasonal variations and geography[J].Tropical Animal Health & Production,2009,41(7):1197-1202.
[2] 赵文华,杨仕标,蒋梅,等.小反刍兽疫病毒的N、M、F及H基因的序列测定和分析[J].动物医学进展,2007,28(11):8-11.ZHAO W H,YANG S B,JIANG M,et al.Sequencing and analysis of N,M,F and H genes of PPRV[J].Progress in Veterinary Medicine,2007,28(11):8-11.(in Chinese)
[3] 刘永宏,赵丽,曹胜波,等.反刍兽疫在中国的流行趋势及应对措施[J].动物医学进展,2015,36(1):110-113.LIU Y H,ZHAO L,CAO S B,et al.Prevalent trends and counter measures of peste des petits ruminants in China[J].Progress in Veterinary Medicine,2015,36(1):110-113.(in Chinese)
[4] 赵玲娜,金红岩,梁琳,等.实时荧光定量RT-PCR检测小反刍兽疫病毒方法的建立[J].中国畜牧兽医,2016,43(11):2844-2851.ZHAO L N,JIN H Y,LIANG L,et al.A SYBR greenⅠ based on Real-time quantitative RT-PCR assay for specificdetection of peste des petits ruminants virus[J].China Animal Science & Veterinary Medicine,2016,43(11):2844-2851.(in Chinese)
[5] 汪萍,尹传振,柏庆,等.阻断ELISA与病毒中和试验检测PPR抗体的比较分析[J].中国预防兽医学报,2016,38(8):658-660.WANG P,YIN C Z,BAI Q,et al.Comparison of ELISA with the virus neutralization test for detection of peste des petits ruminants antibodies in sera[J].Chinese Journal of Preventive Veterinary Medicine,2016,38(8):658-660.(in Chinese)
[6] 李景玉,高玉竹,李元果,等.小反刍兽疫研究进展[J].中国畜牧兽医,2014,41(10):274-280.LI J Y,GAO Y Z,LI Y G,et al.Research progress on peste des petits ruminants[J].China Animal Science & Veterinary Medicine,2014,41(10):274-280.(in Chinese)
[7] 龙云凤,刘晓慧,周晓黎,等.小反刍兽疫流行病学及防控研究进展[J].动物医学进展,2012,33(5):94-98.LONG Y F,LIU X H,ZHOU X L,et al.Epidemiology of peste des petits ruminants and advances in prevention and control[J].Progress in Veterinary Medicine,2012,33(5):94-98.(in Chinese)
[8] 信爱国,杨仁标,向文彬.小反刍兽疫研究进展[J].中国预防兽医学报,2003,25(2):73-76.XIN A G,YANG R B,XIANG W B.Research progress on peste des petits ruminants[J].Chinese Journal of Preventive Veterinary Medicine,2003,25(2):73-76.(in Chinese)
[9] LANGEDIJK J P,DAUS F J,VAN OIRSCHOT J T.Sequence and structure alignment of Paramyxoviridae attachment proteins and discovery of enzymatic activity for a morbillivirus hemagglutinin[J].Journal of Virology,1997,71(8):6155-6167.
[10] ZHAO L N,JIN H Y,XUE X J,et al.Effect of PPRV H gene overexpression on PPRV multiplication[J].Chinese Veterinary Science,2016,46(11):1349-1357.
[11] 刘玉洪,徐自忠,花群义,等.小反刍兽疫病毒分子生物学研究进展[J].动物医学进展,2006,27(12):1-6.LIU Y H,XU Z Z,HUA Q Y,et al.Advance in molecular biology of peste des petits ruminants virus[J].Progress in Veterinary Medicine,2006,27(12):1-6.(in Chinese)
[12] LI J D,MENG X L,ZHU X L,et al.Study on recombinant goat pox virus vaccines vectored with H or F gene of peste des petits ruminants virus[J].Chinese Veterinary Science,2015,45(2):111-117.
[13] 董浩,段小波.小反刍兽疫分子生物学研究进展[J].中国畜牧兽医,2011,38(10):137-140.DONG H,DUAN X B.Advance in molecular biology of peste des petits ruminants virus[J].China Journal of Animal Husbandry & Veterinary Medicine,2011,38(10):137-140.(in Chinese)
[14] 邱文英,李伟,李刚,等.小反刍兽疫N蛋白单克隆抗体的制备及竞争ELISA方法的建立[J].农业生物技术学报,2011,19(5):967-972.QIU W Y,LI W,LI G,et al.Preparation of monoclonal antibodies against peste des petits ruminant virus (PPRV) nucleoprotein and establishment of a competitive ELISA for the detection of PPRV antibodies[J].Journal of Agricultural Biotechnology,2011,19(5):967-972.(in Chinese)
[15] 刘文华,包静月,吴晓东,等.中国小反刍兽疫病毒分离株China/Tib/07全基因组序列测定与分析[J].病毒学报,2010,26(4):322-329.LIU W H,BAO J Y,WU X D,et al.Genome sequencing and analysis of a peste des petits ruminants virus isolate,China/Tib/07[J].Chinese Journal of Virology,2010,26(4):322-329.(in Chinese)
[16] PANG W,MINGZHE F U,ZHANG Q,et al.Establishment and application of PPRV indirect ELISA detection method based on N protein[J].Journal of Northwest A & F University,2017,45(11):1-8.
[17] 高佳滨,许立军,郭艳芹,等.小反刍兽疫研究进展[J].吉林畜牧兽医,2016,10:9-11.GAO J B,XU L J,GUO Y Q,et al.Research progress on peste des petits ruminant[J]. Jilin Animal Husbandry and Veterinary Medicine,2016,10:9-11.(in Chinese)
[18] POLCI A,COSSEDDU G M,ANCORA M,et al.Development and preliminary evaluation of a new Real-time RT-PCR Assay for detection of peste des petits ruminants virus genome[J].Transboundary & Emerging Diseases,2015,62(3):332.
[19] KWIATEK O,MINET C,GRILLET C,et al.Peste des petits ruminants (PPR) outbreak in Tajikistan[J].Journal of Comparative Pathology,2007,136(2-3):111-119.
[20] 于恒智,刘晔,程玮,等.5种外来动物疫病并联荧光定量PCR检测方法的研究[J].中国动物检疫,2016,33(11):79-85.YU H Z,LIU Y,CHENG W,et al.Development of parallel fluorescene quantitative PCR for detection of the five exotic animal diseases[J]. China Animal Health Inspection,2016,33(11):79-85.(in Chinese)
[21] 王凡,刘建斌,祝秀梅.小反刍兽疫病毒研究进展[J].现代生物医学进展,2009,9(14):2776-2777.WANG F,LIU J B,ZHU X M.Current advances on molecular biology of peste des petits ruminants virus[J].Progress in Modern Biomedicine,2009,9(14):2776-2777.(in Chinese)
[22] 赵亚,马海璐,李岳洋,等.小反刍兽疫病毒新疆株的N基因序列分析[J].畜牧与兽医,2016,48(2):17-23.ZHAO Y,MA H L,LI Y Y,et al.Sequence analysis of N gene of peste des petits ruminants virus from Xinjiang province[J].Animal Husbandry & Veterinary Medicine,2016,48(2):17-23.(in Chinese)
[23] BARON M D.The Molecular Biology of Peste des Petits Ruminants Virus[M].Heidelberg:Springer,2015.
[24] 李伟,李刚,邱文英,等.小反刍兽疫病毒H基因的原核表达与鉴定[J].中国预防兽医学报,2009,31(2):117-121.LI W,LI G,QIU W Y,et al.Prokaryotic expression of peste des petits ruminants virus H gene[J].Chinese Journal of Preventive Veterinary Medicine,2009,31(2):117-121.(in Chinese)
[25] SHAILA M S,SHAMAKI D,FORSYTH M A,et al.Geographic distribution and epidemiology of peste des petits ruminants virus[J].Virus Research,1996,43(2):149-153.

Molecular Characteristics and Cluster of PPRV Guizhou Strains Based on N Gene

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	SONG Chun, WANG Bolin, LI Mei, WEN Ming, WANG Kaigong, CHEN Changxiu, CHENG Zhentao. Isolation and Identification of Mycoplasma synoviae Guizhou Strain and Sequence Analysis of VlhA Gene [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1516-1523.
[2]	WEI Yunfang, LI Feixiang, Xing Yun, Huang Qingguo, LI Judong, WAN Jiusheng, CHEN Fangliang. Study on Cloning, Bioinformatics Analysis and Spatio-temporal Expression of IGF2 Gene in Kunming Dogs [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 866-875.
[3]	HE Xianming, REN Guangcai, YE Junxian, LAN Hong, LIU Yufu, XIONG Ting, YANG Zekun, XU Ting, CHEN Ruiai. Isolation and Identification of a Strain of Infectious Bursal Disease Virus and Sequence Analysis of VP2 Gene [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 1005-1014.
[4]	CAI Jie, WU Jiayi, PAN Yuanxia, HU Gaowei, ZHANG Linzhou. Isolation, Identification and Drug Resistance Analysis of Chryseobacterium sp. from Pseudosciaena crocea [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 1135-1143.
[5]	HU Xiaobing, LIN Biaosheng, WANG Zhenwei, ZHENG Lijing. Isolation, Identification and Genomic Sequence Analysis of a Strain of Clostridium butyricum from Pigs Utilizing Lactic Acid [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 569-578.
[6]	KOU Meiling, MIAO Haisheng, LI Le, XIE Jiarui, LI Huachun, SONG Jianling. Isolation and Identification of Chuzan Virus from Palyam Serogroup Virus in Yunnan Border Area [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4401-4409.
[7]	TAN Bin, WANG Chao, WANG Yan, ZHANG Shuqin. Isolation and Identification of Parvovirus from Palm Civets and Sequence Analysis of VP2 Gene [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(10): 4012-4018.
[8]	WANG Zhiyan, SUN Yujie, XU Xin, HUANG Baicheng, TIAN Kegong. Isolation,Identification and Pathogenicity of Porcine Deltacoronavirus Strain HeN10 [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 2982-2989.
[9]	LI Nan, HE Yuwen, ZUO Yuanyuan, MENG Jinxin, XU Tiangang, WANG Jinglin. Analysis of Molecular Biological Characteristics of Structural Genes of Getah Virus (SZC30) Isolated from Midge [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(4): 1149-1160.
[10]	ZHANG Qi, CHEN Haotian, SHAN Jiabao, CHANG Rui, ZHANG Xue, WANG Yao, WANG Junwei, MA Bo. Cloning and Sequence Analysis of NS1 and VP1 Genes of Gosling Plague Virus from Jilin District [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(4): 1161-1169.
[11]	GUO Mengjiao, LI Mingtao, TAN Huihui, ZHANG Chengcheng, ZHANG Xiaorong, WU Yantao. Study on Cloning and Expression Characteristic of NLRC3 Gene in Rabbit [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(11): 3940-3949.
[12]	WANG Zhengfan, ZHU Lisai, LI Fei, XIANG Rui, LI Yiyun, YING Biyun, WANG Guiping, BAI Aiquan, JIA Aiqing. Pathogenicity and Whole Genome Sequence Analysis of Porcine Deltacoronavirus CH7328 Strain [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(11): 4143-4153.
[13]	YIN Dejing, ZHANG Yundan, YUE Jun, LI Qiaobin, HE Ling, LI Tao, WANG Kaigong, CHENG Zhentao. Whole Genomic Sequencing Analysis of Fowl Adenovirus Serotype 4 from Guizhou [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(1): 44-54.
[14]	PAN Pengcheng, WEN Binhua, XIE Wan, JIANG Changjin, JIAO Di, CHEN Baojian, GUAN Zhihui, XIE Bingkun. Sequence Analysis,Eukaryotic Expression Vector Construction and Tissue Expression Analysis of PDK4 Gene in Luchuan Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(8): 2337-2347.
[15]	QIN Benyuan, YANG Shuai, ZHANG Yanwei, LI Wenxin, LIU Hong, WU Yiqi, ZHANG Xuelian, LI Wenxia, YANG Yang, CAI Chunbo, GAO Pengfei, GUO Xiaohong, LI Bugao, CAO Guoqing. Study on Cloning,Bioinformatics Analysis and Expression Profile of ASB2 Gene CDS in Pig [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(5): 1289-1298.