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Cloning and Phylogenetic Analysis of NS1 Genes from Different Isolates of H9N2 Subtype Duck Influenza Virus
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作者 谢青梅 张祥斌 +3 位作者 吴志强 冀君 周科 毕英佐 《Agricultural Science & Technology》 CAS 2009年第1期64-67,126,共5页
[ Objective] The study aimed to lay a foundation for the further studies on function mechanism of NS1 protein in the interspecies transmission of waterfowl influenza virus. [Method] Using the serologic assay and the s... [ Objective] The study aimed to lay a foundation for the further studies on function mechanism of NS1 protein in the interspecies transmission of waterfowl influenza virus. [Method] Using the serologic assay and the specific RT-PCR method, some strains of H9 subtype waterfowl influenza virus were isolated from the 12 to 20 day-old muscovy duck flocks without any clinical symptoms in different areas of Guangdong Province. Four of these strains, including A/duck/ZQ/303/2007(H9N2) (A3 for short), A/Duck/FJ/301/2007 (H9N2) (C1 for short), A/Duck/NH/306/2007(H9N2) ( D6 for short), A/duck/SS/402/2007(H9N2) ( E2 for short), and a strain named A/duck/ZC/2007(H9N2) (L1 for short) from a muscovy duck died of avian influenza virus (AIV), were used for NSl gene cloning and sequencing. Subsequently, the obtained NSl gene sequences were compared with other NS1 sequences registered in GenBank, and the phylogenetic analysis was also conducted. [Result] When compared with the H9N2 AIV NS1 sequences in GenBank, the NSl genes of the four AIV strains A3, C1, 136 and E2 displayed homologies ranging from 99% to 100% at nucleotide level, and 95% to 100% at amino acid level; while the NSl gene of L1 strain displayed homology ranging from 94% to 97% at nucleotide level, and 93% to 98% at amino acid level. The phylogenetic tree demonstrated that A3, C1, D6 and E2 were highly resemblant, and L1 was closest to AY66473 (chicken, 2003). By comparison with the NS1 gene sequences of L1, AF523514 (duck), AY664743 (chicken) and EF155262.1 (quail) using DNAstar, A3, C1, D6 and E.2 presented nucleotide variations at site 21 ( R→Q), 70, 71 ( KE→EG), 86 ( A→S), 124 (V→M) and 225 ( S→N), and amino acid variations at site 21,70, 71 and 86 in dsRNA- dependent protein kinase (PKR) binding domain of NSl gene, which induced the evident variations of antigenic determinant and surface proba- bility plot of NS1 protein. [ Conclusion] This study suggested that the amino acid sequence variation in PKR binding domain of NS1 protein had something to do with the virus pathogenicity. 展开更多
关键词 H9N2 subtype duck influenza virus NS1 gene PKR Phylogenetic analysis
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Cloning and Sequence of Glycoprotein H Gene of Duck Plague Virus 被引量:12
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作者 HAN Xian-jie WANG Jun-wei MA Bo 《Agricultural Sciences in China》 CAS CSCD 2006年第5期397-402,共6页
The glycoprotein H (gH) gene homologue of duck plague virus (DPV) was cloned by degenerate polymerase chain reaction (PCR) and sequenced. It was located immediately downstream from the thymidine kinase gene (TK... The glycoprotein H (gH) gene homologue of duck plague virus (DPV) was cloned by degenerate polymerase chain reaction (PCR) and sequenced. It was located immediately downstream from the thymidine kinase gene (TK). In addition, the 3'-end of the gene homologue to herpesvirus UL21 was located downstream from the gH gene. DPV gH gene open reading frame (ORF) was 2 505 bp in length and its primary translation product was a polypeptide of 834 amino acids long. It possessed several characteristics of membrane glycoproteins, including an N-terminal hydrophobic signal sequence, an external domain containing eight putative N-linked glycosylation sites, a C-terminal transmembrane domain, and a charged cytoplasmic tail. Comparison with other herpesvirus revealed identities of 20.2, 25.1, 23.0, 23.0, 26.5 and 26.0% with the gH counterparts of the human herpesvirus virus 1 (HSV1), equine herpesvirus 4 (EHV4), bovine herpesvirus 1 (BHV1), pseudorabies virus (PRV), gallid herpesvirus 2 (GHV2) and gallid herpesvirus 3 (GHV3), respectively. 展开更多
关键词 duck plague virus glycoprotein H gene degenerate PCR
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Cloning of Thymidine Kinase Gene of Duck Plague Virus Using Degenerate PCR 被引量:11
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作者 HAN Xian-jie WANG Jun-wei 《Agricultural Sciences in China》 CAS CSCD 2005年第8期634-640,共7页
The DNA of duck plague virus (DPV) thymidine kinase (TK) gene was cloned and sequenced from a vaccine virus in the study. Degenerate oligonucleotide primers for the consensus site of herpesvirus UL24, TK, and glyc... The DNA of duck plague virus (DPV) thymidine kinase (TK) gene was cloned and sequenced from a vaccine virus in the study. Degenerate oligonucleotide primers for the consensus site of herpesvirus UL24, TK, and glycoprotein H(gH) gene were used in the polymerase chain reaction (PCR) to amplify DNA product with 3 741-base-pairs (bp) in size. DNA sequence analysis revealed a 1 077-base-pairs (bp) open reading frame (ORF) encoding a 358 amino acid polypeptide homologous to herpesvirus TK proteins. The predicted TK protein shared 31.2, 41.3, 35.7, 37.4, and 28.4% identity with herpes simplex virus typel, equine herpesvirus type 4, Marek's disease virus 2, herpesvirus turkey, and infectious laryngotracheitis virus, respectively. Comparison of the amino acid sequences of other herpesvirus TK proteins showed that these proteins were not conserved on the whole, otherwise the portion of the TK proteins corresponding to the nucleotide binding domain and the nucleoside binding site were highly conserved among herpesvirus. Comparison with the amino acid sequences of the conserved nucleotide and nucleoside binding domains of other eleven herpesvirus TK proteins to the predicted DPV peptide confirmed its identity as the DPV TK protein. 展开更多
关键词 duck plague virus Degenerate PCR Thymidine kinase gene
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Role of duck plague virus glycoprotein C in viral adsorption:Absence of specific interactions with cell surface heparan sulfate 被引量:5
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作者 JING Yan-chun WU Ying +9 位作者 SUN Kun-feng WANG Ming-shu CHENG An-chun CHEN Shun JIA Ren-yong ZHU De-kang LIU Ma-feng YANG Qiao JING Bo CHEN Xiao-yue 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2017年第5期1145-1152,共8页
Many mammalian herpes viruses utilize heparan sulfate (HS) moieties present on cell surface proteoglycans as receptors for cell entry, and this process also requires viral glycoprotein C (gC) homologues. However, ... Many mammalian herpes viruses utilize heparan sulfate (HS) moieties present on cell surface proteoglycans as receptors for cell entry, and this process also requires viral glycoprotein C (gC) homologues. However, our understanding of the role of gC in facilitating attachment of other alpha-herpes viruses such as the duck plague virus (DPV) remains preliminary. To study the role of gC during DPV infection, we used a gC-deleted mutant virus (DPV-AgC-EGFP). Examination of the viral copy number by real-time PCR, as well as time course studies of viral adsorption and proliferation revealed that gC was involved in the viral binding to the cell surface. The affinity of viral glycoproteins (gB-DPV, gC-DPV, and gE-DPV) to HS was assessed using a prokaryotic expression system and HJTrapTM HeparJn HP column chromatography. In addition, to confirm that gC played a role in the interaction between DPV and HS, viruses were treated with the HS analogue heparin and host cells were treated with its inhibitors heparinase prior to exposure to DPV-△gC-EGFP or wild-type strain Chinese virulent duck plague virus (DPV-CHv). The effects of heparin and heparinase on virus infectivity demonstrated that function of gC on Viral adsorption is independent of interactions between gC and heparin sulfate on cell surface. All in all, this study demonstrated that the gC of DPV can mediate viral adsorption in an HS-independent manner, which distinguish it from the gC of some other alpha-herpes viruses. Future studies will be required to identify the receptors involved in gC protein binding to cells. This work provides us a foundation for further studies of examining the roles of gC in the adsorption during duck plague virus infection. 展开更多
关键词 duck plague virus (DPV) glycoprotein C (gC) heparan sulfate (HS) viral adsorption
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Genetic Variation of the VP1 Gene of the Virulent Duck Hepatitis A Virus Type 1(DHAV-1) Isolates in Shandong Province of China 被引量:13
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作者 Jiming Gao Junhao Chen +5 位作者 Xingkui Si Zhijing Xie Yanli Zhu Xingxiao Zhang Shujing Wang Shijin Jiang 《Virologica Sinica》 CAS CSCD 2012年第4期248-253,共6页
To investigate the relationship of the variation of virulence and the external capsid proteins of the pandemic duck hepatitis A virus type 1(DHAV-1) isolates,the virulence,cross neutralization assays and the complete ... To investigate the relationship of the variation of virulence and the external capsid proteins of the pandemic duck hepatitis A virus type 1(DHAV-1) isolates,the virulence,cross neutralization assays and the complete sequence of the virion protein 1(VP1) gene of nine virulent DHAV-1 strains,which were isolated from infected ducklings with clinical symptoms in Shandong province of China in 2007-2008,were tested.The fifth generation duck embryo allantoic liquids of the 9 isolates were tested on 12-day-old duck embryos and on 7-day-old ducklings for the median embryonal lethal doses(ELD 50 s) and the median lethal doses(LD 50 s),respectively.The results showed that the ELD 50 s of embryonic duck eggs of the 9 DHAV-1 isolates were between 1.9 × 10 6 /mL to 1.44 × 10 7 /mL,while the LD 50 s were 2.39 × 10 5 /mL to 6.15 × 10 6 /mL.Cross-neutralization tests revealed that the 9 DHAV-1 isolates were completely neutralized by the standard serum and the hyperimmune sera against the 9 DHAV-1 isolates,respectively.Compared with other virulent,moderate virulent,attenuated vaccine and mild strains,the VP1 genes of the 9 strains shared 89.8%-99.7% similarity at the nucleotide level and 92.4%-99.6% at amino acid level with other DHAV-1 strains.There were three hypervariable regions at the C-terminus(aa 158-160,180-193 and 205-219) and other variable points in VP1 protein,but which didn't cause virulence of DHAV-1 change. 展开更多
关键词 duck hepatitis A virus type 1 (DHAV-1) Embryonal lethal dose (ELDs0) Lethal dose (LDso) Cross-neutralization tests Virionprotein 1 (VP 1)
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Effects of Duck Tembusu Virus on Serum Biochemical Indexes, Cytokines and Viral Replication of Ducklings
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作者 曹宗喜 谭树义 +3 位作者 贺冬梅 顾丽红 林哲敏 叶保国 《Agricultural Science & Technology》 CAS 2017年第8期1428-1431,1547,共5页
In order to understand the pathogenicity of duck Tembusu virus (DTMUV), it was injected into muscle of 5-d-old Cherry Valley ducklings according to the dosage of 1×104 EID50. Then, the biochemical indexes of du... In order to understand the pathogenicity of duck Tembusu virus (DTMUV), it was injected into muscle of 5-d-old Cherry Valley ducklings according to the dosage of 1×104 EID50. Then, the biochemical indexes of duckling serum samples were determined by kits, and the changes in detoxification, tissue viral load and cytokines were detected by using fluorescence quantitative PCR. The results showed that DTMUV had serious damage to the liver, kidney, heart and muscle of ducklings; DTMUV could proliferate in the liver, spleen, lung and brain; the virus levels in the liver and brain reached the peaks on day 5 after the inoculation and those in the lung and spleen reached the peaks on day 9; the virus content was highest in the brain, liver and spleen; and DTMUV induced the overexpression of IFN-γ, IFN-α, IL-6, IFN-β, IL-1β, TLR-7,IL-2, major histocompatibility complex type I (MHC-I) andmajor histocompatibility complex type II (MHC-II) in the spleen on day 1 and the overexpression of IL-6 and IL-2 in the brain on days 1, 2 and 3. 展开更多
关键词 duck Tembusu virus (DTMUV) Biochemical index Viral load CYTOKINE PATHOGENICITY
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The Effect of Gankang Suppository on Duck Hepatitis B Virus, Serum Biochemistry and Liver Histology in Ducklings 被引量:1
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作者 李晖 田德英 +2 位作者 吴会玲 陈淼 陈安群 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2008年第4期421-425,共5页
To examine the effect of Gankang Suppository on duck hepatitis B virus (DHBV), the serum biochemistry and hepatic histology in an animal model of DHBV infection, a model of DHBV infection was established by infectin... To examine the effect of Gankang Suppository on duck hepatitis B virus (DHBV), the serum biochemistry and hepatic histology in an animal model of DHBV infection, a model of DHBV infection was established by infecting 1-day-old Yingtaogu ducklings with DHBV-positive serum. The successful model was confirmed by PCR assay and 48 ducklings infected with DHBV were randomly divided into 3 groups: a Gankang Suppository treatment group, an acyclovir (ACV) group and a DHBV model group (control), with each group having 16 animals. All the animals were given the medicines for 4 weeks in a row. The serum of the animals was taken 14 and 28 days after the medica- tion and 7 days after drug discontinuation. Real-time PCR was performed to detect the copy numbers of DHBV DNA in the serum. ALT and AST were dynamically monitored. The ducklings were sacrificed on the 7th day after the discontinuation of the treatment and livers were harvested and examined for inflammation and degeneration of liver cells by using HE staining. The results showed that on day 14, 28 after the treatment and day 7 after the withdrawal, the logarithmic values (log) of DHBV DNA copy numbers in ducklings of Gankang Suppository treatment group were significantly lower than that before the treatment (P=0.0092, P=0.0070, P=0.0080, respectively). Compared with DHBV model control group, the ALT level was significantly decreased (P=0.0020, P=0.0019, respectively) on day 28 after the treatment and on day 7 after the withdrawal. The AST level was also reduced on day 14 after the treatment (P=0.0298). Compared with the ACV control group, the level of ALT was lower on day 7 after the withdrawal (P=0.0016). Histologically, the hepatocyte swelling, vacuolous degeneration and acidophilic degeneration in Gankang Suppository treatment group were alleviated 7 days after the withdrawal as compared with model control group (P=0.0282, P=0.0084, P=0.0195, respectively). It is concluded that Gankang Suppository can effectively suppress DHBV replication, reduce the levels of serum ALT and AST and improve hepatic histology. 展开更多
关键词 duck hepatitis B virus Gankang Suppository duck hepatitis animal model BIOCHEMISTRY HISTOLOGY
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Replication of hepatitis B virus in primary duck hepatocytes transfected with linear viral DNA 被引量:2
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作者 Yun-Qing Yao Ding-Feng Zhang +10 位作者 Ni Tang Ai-Long Huang Xiao-Yi Zou Jiang-Feng Xiao Yun Luo Da-Zhi Zhang Bo Wang Wei-Ping Zhou Hong Ren Qi Liu Shu-Hua Guo 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第32期5019-5021,共3页
AIM: To explore the expression and replication of hepatitis B virus (HBV) DNA in primary duck hepatocytes (PDHs).METHODS: Complete HBV genome was transfected into PDHs by electroporation (transfected group, 1.19×... AIM: To explore the expression and replication of hepatitis B virus (HBV) DNA in primary duck hepatocytes (PDHs).METHODS: Complete HBV genome was transfected into PDHs by electroporation (transfected group, 1.19×1012copies of linear HBV DNA/1×107 PDHs). After 1-5 d of transfection, HBsAg and HBeAg in the supernatant and lysate of PDHs were measured with the IMX System.Meanwhile, replicative intermediates of HBV DNA were analyzed by Southern blotting and Dot blotting. PDHs electroporated were used as control group.RESULTS: HBsAg in the hepatocyte lysates of transfected group was 15.24 (1 d), 14.55 (3 d) and 5.13 (5 d; P/N values, positive≥2.1) respectively. HBeAg was negative (<2.1). Both HBsAg and HBeAg were negative in the supernatant of transfected group. Dot blotting revealed that HBV DNA was strongly positive in the transfected group and negative in the control group. Southern blot analysis of intracellular total DNA indicated that there were relaxed circular (rc DNA), covalently closed circular (ccc DNA), and single-stranded (ss DNA) HBV DNA replicative intermediates in the transfected group, there was no integrated HBV DNA in the cellular genome. These parameters were negative in control group.CONCLUSION: Expression and replication of HBV genes can occur in hepatocytes from non-mammalian species.HBV replication has no critical species-specificity, and yet hepatic-specific regulating factors in hepatocytes may be essential for viral replication. 展开更多
关键词 Hepatitis B virus REPLICATION EXPRESSION Primary duck hepatocytes
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Isolation,Identification and Pathogenic Characteristics of Duck Hepatitis Virus Isolates 被引量:1
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作者 GAO Ya-dong LING Hong-li DONG Rui-e SUN Hai-xin 《Animal Husbandry and Feed Science》 CAS 2010年第8期37-39,共3页
[ Objective] The aim of this study was to determine and analyze the pathogen and the reasons of duck viral hepatitis which is prevalent recently and difficult to control. [Method] Viruses were isolated from livers and... [ Objective] The aim of this study was to determine and analyze the pathogen and the reasons of duck viral hepatitis which is prevalent recently and difficult to control. [Method] Viruses were isolated from livers and spleens of ducks with typical clinical symptoms in Linyi, Weifang, Binzhou and other regions of Shandong Province. The pathogenic characteristics were observed by inoculation in chicken or duck embryo, RT- PCR, serological test, and duck regression. [ Result] Four duck hepatitis virus (DHV) strains were isolated, and the 5th passage allantoic fluid contained 10^3.41 -10^5.20 ELD50/mI. The serum cross protection rate was 20% -80% between the DHV stains and DHV type I. The mortalities of 4- day-old healthy ducks challenged by these four stains were 50% -100%. All challenged ducks had typical lesions of duck viral hepatitis, and the death peak appeared after 24-48 h. [Conclusion] The virulence of different DHV isolates has regional difference. 展开更多
关键词 duck hepatitis virus IDENTIFICATION ISOLATION viruLENCE
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Molecular Characterization and SYBR Green I-Based Quantitative PCR for Duck Hepatitis Virus Type 1 被引量:1
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作者 LUO Yu-jun ZHANG Gui-hong +2 位作者 XU Xiao-qin CHEN Jian-hong LIAO Ming 《Agricultural Sciences in China》 CAS CSCD 2008年第9期1140-1146,共7页
To determine the genomic sequence of a duck hepatitis virus type 1 (DHV-1) strain, real-time quantitative polymerase chain reaction (RTQ-PCR) assay based on SYBR Green I technology was developed to target 3D gene ... To determine the genomic sequence of a duck hepatitis virus type 1 (DHV-1) strain, real-time quantitative polymerase chain reaction (RTQ-PCR) assay based on SYBR Green I technology was developed to target 3D gene of DHV-1, Comparative sequence analysis showed that the genome has a typical picornarivus genetic organization, and strain DHV-1 R genetic organaiztion is 5' untranslated region (UTR)-VP0-VP3-VPI-2A1-2A2-2B-2C-3A-3B-3C-3D-3' UTR, DHV-1 R has close relationship with Parechovirus, and has 95.1-99.1% nucleotide sequence identity with other DHV-1 strains. Based on the DHV-1 sequences in GenBank, three pairs of specific primers were designed to amplify DHV-1 using real-time PCR. The results showed that real-time PCR Tm value is 85.6℃ and the real-time PCR provides a broad dynamic range, detecting from 102 to 109 copies of DHV-1 cDNA per reaction. No cross-reactions were found in specimens containing DPV, AIV and NDV. It is concluded that DHV-1 belongs to a new group of the family Picornaviridae that may form a separate genus most closely related to the genus Parechovirus. All results showed that the real-time PCR has high sensitivity and specificity to detect DHV-1 using SYBR Green I dissociation curve analysis, isolates can be distinguished by their melting temperature. These methods are rapid, sensitive, and reliable, and can be readily adapted for detection of DHV-1 from other clinical samples. 展开更多
关键词 duck hepatitis virus type 1 complete genome real-time RT-PCR
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Replication and Pathology of Duck Influenza Virus Subtype H9N2 in Chukar 被引量:3
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作者 ZHU Yin Chuan ZHANG Bin +6 位作者 SUN Zeng Hu WANG Xi Jing FAN Xiao Hui GAO Ling Xi LIANG Ying CHEN Xiao Yan ZHANG Zeng Feng 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 2018年第4期306-310,共5页
To investigate the susceptibility of Chukars to duck avian influenza virus H9N2 and explore their role in interspecies transmission of influenza viruses.Chukars were inoculated with duck avian influenza viruses H9N2.
关键词 In ST Replication and Pathology of duck Influenza virus Subtype H9N2 in Chukar
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Molecular Characterization of the Duck Enteritis Virus UL4 Gene 被引量:1
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作者 Hua-qi PAN Nan WANG +5 位作者 Li LIU Lei LIU Jiang-chun HU Pu-yan CHEN Shu-jin WANG Rui-bing CAO 《Virologica Sinica》 SCIE CAS CSCD 2009年第3期171-178,共8页
Duck enteritis virus(DEV) is a herpesvirus that causes an acute,contagious and fatal disease. In the present article,the DEV UL4 gene was cloned and sequenced from a vaccine virus. A degenerate oligonucleotide primer ... Duck enteritis virus(DEV) is a herpesvirus that causes an acute,contagious and fatal disease. In the present article,the DEV UL4 gene was cloned and sequenced from a vaccine virus. A degenerate oligonucleotide primer for the consensus site of herpesvirus UL3 gene and a specific primer located in UL5 were used in the polymerase chain reaction(PCR) to amplify a DNA product 2 086 bp in size. DNA sequence analysis revealed that a 714 bp open reading frame(ORF) of DEV encoding a 237 amino acid polypeptide is homologous to the family of herpesvirus UL4 proteins and therefore has been characterized as a DEV UL4 gene. Alignment of the DEV UL4 protein sequence with those of other alphaherpesviruses showed that 10 amino acid residues are completely conserved. Phylogenetic tree analysis showed that the seventeen alphaherpesviruses viruses analyzed were classified into four large groups,and the duck enteritis virus branched separately,closely related to the Mardiviruses group comprising Gallid herpesvirus 2(GaHV-2) ,Gallid herpesvirus 3(GaHV-3) and Meleagrid herpesvirus 1(MeHV-1) . The present study showed that the evolutionary relationship of the UL4 protein could be used for classification of alphaherpesviruses. 展开更多
关键词 duck enteritis virus (DEV) Degenerate primer PCR UL4 gene Gene Character
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Complete Genomic Sequence of a Chinese Isolate of Duck Hepatitis Virus 被引量:9
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作者 Guang-qing LIU Fei WANG +4 位作者 Zheng NI Tao YUN Bin YU Jiong-gang HUANG Jian-Ping CHEN 《中国病毒学》 CSCD 2007年第5期353-359,共7页
The complete genomic sequence of Duck hepatitis virus 1(DHV-1) ZJ-V isolate was sequenced and determined to be 7 691 nucleotides(nt) in length with a 5'-terminal un-translated region(UTR) of 626 nt and a 3'-te... The complete genomic sequence of Duck hepatitis virus 1(DHV-1) ZJ-V isolate was sequenced and determined to be 7 691 nucleotides(nt) in length with a 5'-terminal un-translated region(UTR) of 626 nt and a 3'-terminal UTR of 315 nt(not including the poly(A) tail).One large open reading frame(ORF) was found within the genome(nt 627 to 7 373) coding for a polypeptide of 2 249 amino acids.Our data also showed that the poly(A) tail of DHV-1 has at least 22 A's.Sequence comparison revealed significant homology(from 91.9% to 95.7%) between the protein sequences of the ZJ-V isolate and those of 21 reference isolates.Although DHV-1 has been classified as an unassigned virus in the Picornaviridae family,its genome showed some unique characteristics.DHV-1 contains 3 copies of the 2A gene and only 1 copy of the 3B gene,and its 3'-NCR is longer than those of other picornaviruses.Phylogenetic analysis to do sequence homology based on the VP1 protein sequences showed that the ZJ-V isolate shares high sequence homology with the reported DHV-1 isolates(from 92.9% to 99.2%),indicating that DHV-1 is genetically stable. 展开更多
关键词 全基因序列 鸭肝炎病毒 中国 隔离群
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The codon-optimized capsid gene of duck circovirus can be highly expressed in yeast and self-assemble into virus-like particles 被引量:1
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作者 YANG Cui XU Yu +10 位作者 JIA Ren-yong LIU Si-yang WANG Ming-shu ZHU De-kang CHEN Shun LIU Ma-feng ZHAO Xin-xin SUN Kun-feng JING Bo YIN Zhong-qiong CHENG An-chun 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2017年第7期1601-1608,共8页
The capsid (Cap) protein, which is the only structural protein of duck circovirus (DuCV), is the most important antigen for the development of vaccines against DuCV and the virus's serological diagnostic methods.... The capsid (Cap) protein, which is the only structural protein of duck circovirus (DuCV), is the most important antigen for the development of vaccines against DuCV and the virus's serological diagnostic methods. In order to use yeast expression system to produce a large quantities of DuCVCap protein which is close to its natural form to display the antigen peptides perfectly, the Cap gene was optimized into the codon-optimized capsid (Opt-Cap) gene towards the preference of yeast firstly. Then, the genes of Cap and Opt-Cap were separately cloned into pPIC9K plasmid and transformed into Picha pas- toris GSl15. The strains that displayed the phenotype of Mut~ and contained multiple inserts of expression cassette were selected from those colonies. After the induction expression, the secretory type of Cap protein, which was about 43 kDa, was best expressed under 0.5% (v/v) methanol and sorbitol induction. Compared with the Cap gene, the expression level of Opt-Cap gene was much higher. What's more, the purified Cap protein had a good reactivity to its specific polyclone antibody and DuCV-positive serum, and it was able to self-assemble into virus-like particles (VLPs). These VLPs, with a diameter of 15-20 nm and without a nucleic acid structure, showed a high level of similarity to DuCV particles in size and shape. All of the resultsdemonstrated that, based on the codon-optimization, it is suitable to use the P. pastoris expression system to produce DuCV VLPs on a large scale. It is the first time that a large amounts of DuCV VLPs were produced successfully in P. pastoris, which might be particularly useful for the further studies of serological diagnosis and vaccines of DuCV. 展开更多
关键词 capsid gene codon'optimization duck circovirus virus-like particles
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The Potential of Duck Hepatitis Virus(DHV-1) Stimulating the Body Weight Gain and the Effects of Silymarin on It in Duckling 被引量:1
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作者 LIU Wei-min WANG Bing-yun CHEN Jian-hong WANG Jun JI Hui-qin YUAN Sheng HUANG De-chun LI Kang-lin 《Agricultural Sciences in China》 CAS CSCD 2009年第11期1403-1408,共6页
To evaluate the effects of duck hepatitis virus-1 (DHV-1) on the body weight gain in duck and the effects of silymarin on it in vivo, 100 10-d-old ducks, both male and female, were collected to be subjected to the t... To evaluate the effects of duck hepatitis virus-1 (DHV-1) on the body weight gain in duck and the effects of silymarin on it in vivo, 100 10-d-old ducks, both male and female, were collected to be subjected to the test. The experiments were conducted in 8 groups: in group 1-3, the animals were inoculated with 1:105 diluted duck hepatitis virus (DHV-1) infected allantoic fluid and given 0, 30, and 50 mg kg^-1 BW d^-1 silymarin orally, respectively. In group 4-6, the animals were inoculated with 1:5 × 105 diluted DHV-1 infected allantoic fluid and given 0, 10, and 30 mg kg^-1 BW d^-1 silymarin orally, respectively. In group 7, the animals were given 10 mg kg^-1 BW d^-1 silymafin only. Group 8 was the control one treated by injecting sterillized saline into the leg muscles. All the silymarin was given from 0 to 4 d after inoculation of the virus. By the 5th d after inoculation, the vein blood was drawn from the dorsal foot vein and the plasma samples were collected and stored at -20℃. The body weight gain (BWG) was measured from 0 to 10 d after inoculation. The plasma IGF-I, T3, and T4 concentrations were measured by radioimmunoassay (RIA). At the virus dose of 1:5 ×105 diluted virus infected allantoic fluid, the inoculation of the virus enhanced the BWG significantly compared with that of the control (P〈 0.01), while 10-50 mg kg^-1 BW d^-1 silymarin could counteract the effects of the virus on the BWG dose-dependently. The plasma IGF-I levels showed no correlation with the BWG, but the T3 levels showed a same tropism with the body weight gain. The present results indicated that sublethal DHV-1 enhanced the body weight gain of ducklings significantly, and the silymarin could counteract this effect in vivo. 展开更多
关键词 duck hepatitis virus-l DHV body weight gain SILYMARIN INFECTION duckLING
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Cloning and sequence analysis of US1 gene in duck enteritis virus 被引量:1
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作者 ZHAO Yan WANG Jun-wei +1 位作者 MA Bo ZHAO Xiao-yan 《中国预防兽医学报》 CAS CSCD 北大核心 2011年第2期105-109,共5页
In this paper,a 1,860 bp sequence in IRs region of duck enteritis virus(DEV) was amplified by single oligonucleotide nested PCR with a single primer designed according to partial sequence of US1 and then a pair of pri... In this paper,a 1,860 bp sequence in IRs region of duck enteritis virus(DEV) was amplified by single oligonucleotide nested PCR with a single primer designed according to partial sequence of US1 and then a pair of primers designed according to the 3' UTR of US8 gene and 5' end of the new getting sequence were used to amplify a 2,426 bp sequence toward the TRs region.Sequence analysis revealed that the both sequences contained an identical 990 bp open reading frame of DEV US1 gene.The two ORFs were in opposite transcription orientation.Sequence comparison of the nucleotide sequence and the deduced amino acid sequence of US1 gene showed relatively high identity to Mardivirus.Phylogenetic tree analysis showed that the eleven herpesviruses viruses were classified into three groups,and the duck enteritis virus was most closely related to Mardivirus. 展开更多
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Identification of novel genes associated with duck OASL in response to influenza A virus
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作者 WANG Xiao-xue LU Chang +6 位作者 RONG En-guang HU Jia-xiang XING Yan-ling LIU Zheng-yu GAO Chu-ze LIU Jin-hua HUANG Yin-hua 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2019年第7期1451-1459,共9页
2′-5′-Oligoadenylate synthetase like protein(OASL) plays a key role in response to viral infections through selectively activating the OAS/RNase L or OASL/RIG-I signaling pathway.Although classic pathway of OASL is ... 2′-5′-Oligoadenylate synthetase like protein(OASL) plays a key role in response to viral infections through selectively activating the OAS/RNase L or OASL/RIG-I signaling pathway.Although classic pathway of OASL is well-known,its regulated genes or co-actors are largely unknown.To study the possible molecular mechanism of duck OASL(dOASL),we performed RNA-sequencing(RNA-seq) and immunoprecipitation and mass spectrometry(IP-MS) at the level of mRNA and protein,respectively.For RNA-seq,we used DF1 cell lines(DF1 dO ASL+/+,DF1 cO ASL–/–,and DF1) with or without the CK/0513 H5 N1 virus(A/chicken/huabei/0513/2007) infection.1 737 differentially expressed genes(DEGs) were identified as candidate target genes regulated by dOASL.Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG) analysis and Weighted Correlation Network Analysis(WGCNA) were performed.We identified one important yellow co-expression module correlated with antiviral immune response.In this module,Ankyrin repeat and FYVE domain containing 1(ANKFY1),harboring a BTB domain similar to the methyl CpG-binding protein 1(MBD1) which bound to OASL in human,was regulated by dOASL.At protein level,133 host proteins were detected.Interestingly,ANKFY1 was one of them binding to dOASL protein.Further phylogenomic and chromosomal syntenic analysis demonstrated MBD1 was absent in birds,while mammals retained.It is suggested that OASL-ANKFY1 interaction might act as a compensatory mechanism to regulate gene expression in birds.Our findings will provide a useful resource for the molecular mechanism research of dOASL. 展开更多
关键词 duck OASL ANKFY1 COMPENSATORY MECHANISM INFLUENZA A virus
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Molecular Characterization of Duck Hepatitis B Virus Isolated from Hubei Brown Ducks
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作者 胡权 张小勇 +3 位作者 雷延昌 张正茂 Mengji Lu 杨东亮 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2006年第5期633-636,共4页
The objective of this study was to characterize the genome structure of duck hepatitis B virus (DHBV) isolated from Hubei brown ducks. The natural carrier rate of DHBV in adult ducks from Hubei area was investigated... The objective of this study was to characterize the genome structure of duck hepatitis B virus (DHBV) isolated from Hubei brown ducks. The natural carrier rate of DHBV in adult ducks from Hubei area was investigated and the DHBV DNA-positive serum screened out. The complete genome of a DHBV strain was amplified by polymerase chain reaction (PCR) and cloned into T vector and sequenced. The results showed that the carrier rate of DHBV in Hubei brown ducks was 10 % This strain (GenBank accession number DQ276978) had a genome of 3024 nucleotides with three overlapping open reading frames encoding the surface, core and polymerase proteins respectively. Comparison of the strain with 17 DHBV strains registered in GenBank revealed a homology from 89.3 % to 93.5 % at the nucleotide level. The sequences of the structural and functional domains of these proteins were highly conserved. The strain was found to share more signature amino acids in the polymerase genes with the "Chinese" DHBV strains than those of the "Western" country strains. This finding was also corroborated by a phylogenetic tree analysis. Therefore, the DQ276978 might belong to a subtype of the Chinese DHBV strains. 展开更多
关键词 duck hepatitis B virus homology analysis GENOME CLONING
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Isolation and PCR Identification of Duck Tembusu Virus
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作者 Huang Lili He Dongmei +5 位作者 Ye Baoguo Zhang Yan Yang Shaoxiong Chen Chaoxi Lin Zhemin Cao Zongxi 《Animal Husbandry and Feed Science》 CAS 2017年第5期318-319,共2页
Duck tembusu virus disease is one of the most serious infectious diseases endangering duck industry. A strain of virus was isolated from a dead duck,and performed PCR identification and sequence analysis. The results ... Duck tembusu virus disease is one of the most serious infectious diseases endangering duck industry. A strain of virus was isolated from a dead duck,and performed PCR identification and sequence analysis. The results showed that the sequence of the isolate shared above 99% homology with duck tembusu virus( DTMUV) sequence on Gen Bank. The result indicated that the isolated virus was DTMUV. 展开更多
关键词 duck Tembusu virus (DTMUV) ISOLATION PCR IDENTIFICATION
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Development of RT PCR Assay for Detection of Duck Hepatitis Virus Type 1
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作者 Wang Yongjuan Cui Pingfu +1 位作者 Liu Bo Meng Ting 《Animal Husbandry and Feed Science》 CAS 2017年第5期308-310,共3页
In this study,a RT-PCR assay for rapid detection of duck hepatitis virus type 1( DHV-1) was established in the presence of a pair of primers designed based on vp1 gene sequence,using the genomic RNA of DHV-1 or other ... In this study,a RT-PCR assay for rapid detection of duck hepatitis virus type 1( DHV-1) was established in the presence of a pair of primers designed based on vp1 gene sequence,using the genomic RNA of DHV-1 or other common viruses as the template. The template was serially diluted 10-fold to determine the reaction sensitivity. Finally,seven liver samples of ducks with suspected DHV-1 infection,which were collected from different regions of Jiangsu Province,were detected with this method. The results showed that a DNA fragment of about 360 bp was specifically amplified with the RT-PCR system,and the detection limit was1 fg/μL. The detection rate of clinical samples with this method was 100%. The results revealed that the RT-PCR system which had high specificity and high sensitivity in the detection of DHV-1 was successfully established. 展开更多
关键词 duck HEPATITIS virus TYPE 1 RT PCR DETECTION
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