Characterization of the Duck Plague Virus UL35 Gene

Cai, Mingsheng; Cheng, Anchun; Wang, Mingshu; Chen, Wanping; Zhang, Xian; Zheng, Shang-Xi; Pu, Yang; Lou, Kunpeng; Zhang, Yao; Sun, Lei; Wang, Lulu; Zhu, Dekang; Luo, Qihui; Chen, Xiaoyue

doi:10.1159/000317291

Cited by 14 publications

(7 citation statements)

References 71 publications

(44 reference statements)

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“…IFA was conducted using a standard procedure [ 36 ]. Briefly, DEV-infected DEF cells were plated onto coverslips and fixed with 4 % paraformaldehyde for 30 min at 0, 2, 4, 6, 8, 12, 24, 36, 48, 60 and 72 h post-infection.…”

Section: Methodsmentioning

confidence: 99%

Duck enteritis virus UL54 is an IE protein primarily located in the nucleus

Liu

Cheng

Wang

et al. 2015

Virol J

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BackgroundThe UL54 protein of Duck Enteritis Virus (DEV) is a homolog of herpes simplex virus-1 (HSV-1) immediate-early infectious cell protein 27 (ICP27), a multifunctional protein essential for viral infection. Nonetheless, there is little information on the UL54 protein of DEV.MethodsThe UL54 gene was cloned into the pPAL7 vector, and the recombinant protein, expressed in the E. coli Rosetta, was used to produce a specific antibody. Using this antibody, Western blotting and indirect immunofluorescence analysis (IFA) were used to analyze the expression level and intracellular localization, respectively, of UL54 in DEV-infected cells at different times. Real-time quantitative reverse transcription PCR (RT-PCR) and the pharmacological inhibition test were utilized to ascertain the kinetic class of the UL54 gene.ResultsUL54 was expressed as a fusion protein of approximately 66.0 kDa using the prokaryotic expression system, and this protein was used to generate the specific anti-UL54 antibody. The UL54 protein was initially diffusely distributed throughout the cytoplasmic region; then, after 2 h, it gradually distributed into the nucleus, peaking at 24 h, and complete localization to the nucleus was observed thereafter. The UL54 transcript was detected as early as 0.5 h, and peak expression was observed at 24 h. The UL54 gene was insensitive to the DNA polymerase inhibitor Ganciclovir (GCV) and the protein synthesis inhibitor Cycloheximide (CHX), both of which confirmed that UL54 was an immediate early gene.ConclusionsThe DEV UL54 gene was expressed in a prokaryotic expression system and characterized for expression level, intracellular localization and gene kinetic class. We propose that these results will provide the foundation for further functional analyses of this gene.

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Section: Methodsmentioning

confidence: 99%

Duck enteritis virus UL54 is an IE protein primarily located in the nucleus

Liu

Cheng

Wang

et al. 2015

Virol J

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“…In 1923 the first known occurrence of DVE was recorded in the Netherlands (Mondal et al, 2010, Baudet, 1923.The first DP Virus (DPV) isolation and identification study was published in Bangladesh in 1982, subsequently evaluating the immunogenicity of the DPV vaccine from local isolates (Wu et al, 2012a, Ahamed et al, 2015a, Hossain et al, 2005a, Islam et al, 2005. The morbidity and mortality rates of suspected birds vary between 5% and 100%, depending on the virulence of bird infection and immunological status (Neher et al, 2019) that causes acute hemorrhagic, septic, infectious, and fatal diseases among waterfowls such as ducks, geese, and swans, (Li et al, 2009, You et al, 2018, El-Tholoth et al, 2019, Wu et al, 2012a, which caused major economic losses in the commercial waterfowl industry around the world as a result of sudden deaths, condemnations, and reduced production of eggs (Cai et al, 2010, Lian et al, 2010. Natural infections of waterfowls are susceptible at all ages and there reported that in young ducks from 7 days of age (El-Samadony et al, 2013, El-Tholoth et al, 2019) and infected with direct or indirect contact and developed clinical manifestation within 3 to 7 days after infection (Wang et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

Molecular Characterization of Duck Plague Virus for Determination of TCID<sub>50</sub>

Jahan

Rahman

Ahmed

et al. 2021

Res. Agric. Livest. Fish.

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Duck plague is an enveloped DNA virus that belongs to the Anatid Herpes Virus; the Herpesviridae family is an acute and highly infectious duck, geese, and swan disease that causes tremendous economic losses of duck rearing in Bangladesh and other duck rearing countries. Therefore, we decided to isolate duck plague virus from recent fields’ outbreaks area and performed molecular detection and phylogenetic analysis to find out the similarities between our findings and other isolates around the world. Visceral organs of 13 suspected ducks from recent outbreaks area were collected by post-mortem examination for inoculum preparation. Several passages were performed to harvest into 9-11 old embryonated eggs Chorioallantoic membrane (CAM) route and duck embryo fibroblast (DEF) primary cell culture. DNA polymerase (446bp) and DNA polymerase (UL, 602bp) genes were used for molecular detection by Polymerase chain reaction (PCR). Pathogenicity was done with duckling and TCID50 on DEF. Molecular characterization was performed from extracted DNA of duckling and 2 Positive PCR products were partially sequenced for phylogenetic analysis of their origin and nucleotide variations. Sequenced data was analyzed to reveal genetic relationships among constructed phylogenetic tree for understanding potential transmission with origin of virus and data was then submitted to gene bank and got accession number for DPV-BR1-MN937272 and DPV-BR-2-MN937273. Among 13 samples, 4(30.77%) were found positive by PCR using DNA polymerase at 446 bp and UL at 602 bp gene. Chorioallantoic membrane (CAM) was observed hemorrhagic after 72 days and duck embryo fibroblast (DEF) become round as showed cytopathic characteristics after 48h of infection .Duckling showed that isolated virus was highly pathogenic as characteristics signs of post-mortem examination. Therefore, this has found that recent isolates have similarity with Bangladesh, India and China isolates. Moreover, TCID50 has confirmed the isolates have accepted titer to be a vaccine strain. Res. Agric., Livest. Fish.8(1): 125-133, April 2021

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“…VP26 is the smallest capsid protein in herpes simplex virus type 1 [7] , the ORF of DEV UL35 encodes VP26 comprising 117 amino acids, with molecular mass of 12.9 kDa. DEV VP26 was testi ed to be a component of extracellular mature DEV virions [8] .…”

Section: Introductionmentioning

confidence: 99%

Simultaneous Tracking of Capsid, Envelope Protein Localization in Living Cells Infected with Double Fluorescent Duck Enteritis Virus

Chen

Zheng

Hua

et al. 2020

Preprint

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Background Duck enteritis virus can cause an acute, contagious and lethal disease of many species of waterfowl. An infectious bacterial artificial chromosome clone of duck enteritis virus (DEV) vaccine strain pDEV-EF1 has been constructed in our previous study. To visually studying DEV, a double-labeled recombinant virus have been constructed. Methods based on pDEV-EF1, a recombinant mutated clone pDEV-UL35(c)CFP-gC(c)mRFP which carries a red fluorescent protein(mRFP) gene attached to the viral envelope protein gC, combined with a cyan fluorescent protein (CFP) gene fused to the viral capsid UL35 gene was constructed by two-step Red/ET recombination and the recombinant virus rDEV-UL35(c)CFP-gC(c)mRFP was rescued from chicken embryo fibroblasts (CEFs) by calcium phosphate precipitation. Then protein expression were detected by Western blot analysis, and subcellular location of gC and UL35 were observed by confocal microscopy. Viral morphogenesis were observed by transmission electron microscopy (TEM). Results The recombinant virus rDEV-UL35(c)CFP-gC(c)mRFP was rescued from chicken embryo fibroblasts (CEFs) by calcium phosphate precipitation. Western blot analysis showed UL35-CFP and gC-mRFP are both expressed in fusion forms in rDEV-UL35(c)CFP-gC(c)mRFP-infected CEFs, and subcellular location study showed gC-mRFP was mainly localized in whole cell at 36 h.p.i. and 48 h.p.i.; and then mostly migrated to cytoplasm after 60 h.p.i.; UL35-CFP was localized in nucleus in all stages of virus infection. Transmission electron microscopy indicated that viral particles at different stages of morphogenesis (A capsids, B capsids, C capsids) were observed in virus-infected cells. However,mature C capsids was less in rDEV-UL35(c)CFP-gC(c)mRFP-infected cells than rDEV-dEF1GFP and rDEV-gC(c)mRFP-infected samples. Conclusions This study has laid a foundation for visually studying localization, transportation of DEV capsid protein and envelope glycoprotein, as well as virus assembly, virion movement and virus-cell interaction.

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Characterization of the Duck Plague Virus UL35 Gene

Cited by 14 publications

References 71 publications

Duck enteritis virus UL54 is an IE protein primarily located in the nucleus

Duck enteritis virus UL54 is an IE protein primarily located in the nucleus

Molecular Characterization of Duck Plague Virus for Determination of TCID<sub>50</sub>

Simultaneous Tracking of Capsid, Envelope Protein Localization in Living Cells Infected with Double Fluorescent Duck Enteritis Virus

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