1.Research progress in active substances and their mechanisms of action against porcine epidemic diarrhea virus.
Yu LIU ; Sisi SU ; Ziqian WANG ; Jiahao WU ; Hongwei CHEN ; Hongzao YANG
Chinese Journal of Biotechnology 2025;41(7):2519-2533
Porcine epidemic diarrhea virus (PEDV) is an intestinal coronavirus that can cause porcine epidemic diarrhea, leading to diarrhea, vomiting, weight loss, and even death in piglets. Due to the diversity of PEDV strains, traditional vaccines are difficult to sustainably and effectively prevent and control PEDV. This article reviews the strategies and mechanisms of active substances in regulating intracellular signaling pathways, viral proteins, and microbial metabolites to enhance the host immune function against PEDV. It emphasizes the prevention of PEDV resistance and the potential harm of PEDV breaking through interspecies barriers to the human society, aiming to provide reliable theoretical support for the development of new antiviral drugs or vaccines.
Porcine epidemic diarrhea virus/immunology*
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Animals
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Swine
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Swine Diseases/prevention & control*
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Antiviral Agents/pharmacology*
;
Coronavirus Infections/virology*
;
Viral Vaccines/immunology*
;
Humans
;
Signal Transduction
2.A truncated N protein-based ELISA method for the detection of antibodies against porcine deltacoronavirus.
Dongsheng WANG ; Ruiming YU ; Liping ZHANG ; Yingjie BAI ; Xia LIU ; Yonglu WANG ; Xiaohua DU ; Xinsheng LIU
Chinese Journal of Biotechnology 2025;41(7):2760-2773
This study aims to establish an antibody detection method for porcine deltacoronavirus (PDCoV). The recombinant proteins PDCoV-N1 and PDCoV-N2 were expressed via the prokaryotic plasmid pColdII harboring the N gene sequence of the PDCoV strain CH/XJYN/2016. The reactivity and specificity of PDCoV-N1 and PDCoV-N2 with anti-PEDV sera were analyzed after the recombinant proteins were analyzed by SDS-PAGE and purified by the Ni-NTA Superflow Cartridge. Meanwhile, Western blotting and indirect immunofluorescence assay were carried out separately to validate the recombinant proteins PDCoV-N1 and PDCoV-N2. Finally, we established an indirect ELISA method based on the recombinant protein PDCoV-N2 after optimizing the conditions and tested the sensitivity, specificity, and reproducibility of the method. Then, the established method was employed to examine 102 clinical serum samples. The recombinant protein PDCoV-N2 showed low cross-reactivity with anti-PEDV sera. The optimal conditions of the indirect ELISA method based on PDCoV-N2 were as follows: the antigen coating concentration of 1.25 μg/mL and coating at 37 ℃ for 1 h; blocking by BSA overnight at 4 ℃; serum sample dilution at 1:50 and incubation at 37 ℃ for 1 h; secondary antibody dilution at 1:80 000 and incubation at 37 ℃ for 1 h; color development with TMB chromogenic solution at 37 ℃ for 10 min. The S/P value ≥ 0.45, ≤0.38, and between 0.45 and 0.38 indicated that the test sample was positive, negative, and suspicious, respectively. The testing results of the antisera against porcine epidemic diarrhea virus (PEDV), porcine circovirus 2 (PCV2), transmissible gastroenteritis virus (TGEV), foot-and-mouth disease virus (FMDV), and African swine fever virus (ASFV) showed that the S/P values were all less than 0.38. The testing results of the 800-fold diluted anti-PDCoV sera were still positive. The results of the inter- and intra-batch tests showed that the coefficients of variation of this method were less than 10%. Clinical serum sample test results showed the coincidence rate between this method and neutralization test was 94.12%. In this study, an ELISA method for the detection of anti-PDCoV antibodies was successfully established based on the truncated N protein of PDCoV. This method is sensitive, specific, stable, and reproducible, serving as a new method for the clinical diagnosis of PDCoV.
Animals
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Enzyme-Linked Immunosorbent Assay/methods*
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Swine
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Antibodies, Viral/blood*
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Recombinant Proteins/genetics*
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Deltacoronavirus/isolation & purification*
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Coronavirus Infections/virology*
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Swine Diseases/diagnosis*
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Coronavirus Nucleocapsid Proteins
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Sensitivity and Specificity
3.Nucleocapsid protein from porcine epidemic diarrhea virus isolates can antagonize interferon-λ production by blocking the nuclear factor-κB nuclear translocation.
Ying SHAN ; Zi-Qi LIU ; Guo-Wei LI ; Cong CHEN ; Hao LUO ; Ya-Jie LIU ; Xun-Hui ZHUO ; Xing-Fen SHI ; Wei-Huan FANG ; Xiao-Liang LI
Journal of Zhejiang University. Science. B 2018;19(7):570-580
Porcine epidemic diarrhea virus (PEDV) is a highly infectious pathogen that can cause severe diseases in pigs and result in enormous economic losses in the worldwide swine industry. Previous studies revealed that PEDV exhibits an obvious capacity for modulating interferon (IFN) signaling or expression. The newly discovered type III IFN, which plays a crucial role in antiviral immunity, has strong antiviral activity against PEDV proliferation in IPEC-J2 cells. In this study, we aimed to investigate the effect of PEDV nucleocapsid (N) protein on type III IFN-λ. We found that the N proteins of ten PEDV strains isolated between 2013 and 2017 from different local farms shared high nucleotide identities, while the N protein of the CV777 vaccine strain formed a monophyletic branch in the phylogenetic tree. The N protein of the epidemic strain could antagonize type III IFN, but not type I or type II IFN expression induced by polyinosinic-polycytidylic acid (poly(I:C)) in IPEC-J2 cells. Subsequently, we demonstrated that the inhibition of poly(I:C)-induced IFN-λ3 production by PEDV N protein was dependent on the blocking of nuclear factor-κB (NF-κB) nuclear translocation. These findings might help increase understanding of the pathogenesis of PEDV and its mechanisms for evading the host immune response.
Active Transport, Cell Nucleus
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Animals
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Coronavirus Infections
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immunology
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veterinary
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virology
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Genes, Viral
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Host-Pathogen Interactions
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immunology
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Interferons
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antagonists & inhibitors
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biosynthesis
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genetics
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Interleukins
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antagonists & inhibitors
;
biosynthesis
;
genetics
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NF-kappa B
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metabolism
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Nucleocapsid Proteins
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genetics
;
immunology
;
physiology
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Porcine epidemic diarrhea virus
;
genetics
;
pathogenicity
;
physiology
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Promoter Regions, Genetic
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Swine
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Swine Diseases
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immunology
;
virology
4.Swine-spread severe influenza-associated pneumonia: A case report and literature review.
Ting LIU ; Yafeng JIANG ; Ruoyun OUYANG
Journal of Central South University(Medical Sciences) 2018;43(11):1266-1271
We report and analyze the clinical data of the first case of severe pneumonia caused by influenza B virus from swine. The patient, a 62 year-old male domestic pig breeder, was admitted to hospital because of fever and muscle pain for 5 days, and anhelation for 3 days. One week before the onset of disease, the patient kept close contact with pigs. CT scan of the chest showed diffuse infiltration in both lungs. Influenza B virus antigen detection (colloidal gold method) was repeatedly positive. These all confirmed influenza B virus infection. Poor appetite, weight loss, cough, poor spirit of pigs, positive influenza B virus antigen test occurred in the pig, while the patient had no history of exposure to influenza B-infected patients. It was likely that influenza B virus was transmitted from domestic pigs to the patient by droplets or close contact. Influenza B virus epidemics always occur every five or six years a time, and patients and carriers are the main source of infection. After searching the Pubmed, Web of science, Elsevier, Wanfang, and CNKI databases, it was found that although there were many studies on influenza B virus infecting seals, ferret, domestic pigs, guinea pigs, and other animals, there was no case report for animal-to-human infection. It is the first case report of type B influenza virus transmission from domestic pigs to people in the world, which provides a new direction for the research and prevention of influenza B virus.
Animals
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Humans
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Influenza B virus
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Influenza, Human
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complications
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etiology
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virology
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Lung
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virology
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Male
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Middle Aged
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Orthomyxoviridae Infections
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transmission
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Pneumonia
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etiology
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Swine
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Swine Diseases
;
transmission
;
virology
5.Research Advances in the Porcine Deltacoronavirus.
Puxian FANG ; Liurong FANG ; Nan DONG ; Shaobo XIAO
Chinese Journal of Virology 2016;32(2):243-248
The deltacoronavirus is a new member of the subfamily Coronaviridae of the family Coronaviridae. Deltacoronaviruses can infect birds and mammals. Deltacoronaviruses were detected in early 2007 in Asian leopard cats and Chinese ferret badgers. In 2014, porcine deltacoronavirus (PDCoV) infection spread rapidly in the USA. Moreover, cell culture-adapted PDCoV has been obtained from infected piglets. Animal experiments have confirmed that the isolated PDCoV is highly pathogenic and causes severe diarrhea in piglets. Thus, the PDCoV can be considered to be a good model to study the deltacoronavirus. In this review, we discuss the etiology, epidemiology, pathogenicity, culture, and diagnostic methods of the PDCoV.
Animals
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Coronavirus
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classification
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genetics
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isolation & purification
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Coronavirus Infections
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veterinary
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virology
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Diarrhea
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veterinary
;
virology
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Phylogeny
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Swine
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Swine Diseases
;
virology
6.Immune Response of Recombinant Pseudorabies Virus rPRV-VP2 Expressing VP2 Gene of Porcine Parvovirus in Mice.
Pengfei FU ; Xinlong PAN ; Qiao HAN ; Xingwu YANG ; Qianlei ZHU ; Xiaoqing GUO ; Yu ZHANG ; Hongying CHEN
Chinese Journal of Virology 2016;32(2):195-202
In order to develop a combined live vaccine that will be used to prevent against porcine parvovirus (PPV) and Pseudorabies virus (PRV) infection, the VP2 gene of PPV was inserted into the transfer vector plasmid pG to produce the recombinant plasmid pGVP2. The plasmid pGVP2 and the genome of PRV HB98 attenuated vaccine were transfected by using lipofectamine into swine testis cells for the homologous recombination. The recombinant virus rPRV-VP2 was purified by selection of green fluorescence plaques for five cycles. 6-week-old female Kunming mice were immunized intramuscularly with attenuated PRV parent HB98 strain, commercial inactivated vaccine against PPV, recombinant virus, DMEM culture solution. The injections were repeated with an equivalent dose after 2 weeks in all of the groups, and then challenged with the virulent PRV NY strain at 7 weeks after the first immunization. The recombinant virus rPRV-VP2 was successfully generated, and the recombinant virus could effectively elicite anti-PPV and PRV antibody and significant cellular immune response as indicated by anti-PPV ELISA and HI, PRV-neutralizing assay and flow cytometry. The challenge assay indicated that recombinant virus could protect the mice against the virulent PRV challenge. These results demonstrated that the recombinant virus can be a candidate recombinant vaccine strain for the prevention of PRV and PPV.
Animals
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Antibodies, Viral
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immunology
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Antigens, Viral
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administration & dosage
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genetics
;
immunology
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Capsid Proteins
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administration & dosage
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genetics
;
immunology
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Female
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Gene Expression
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Genetic Vectors
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genetics
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metabolism
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Herpesvirus 1, Suid
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genetics
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metabolism
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Mice
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Parvovirus, Porcine
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genetics
;
immunology
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Swine
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Swine Diseases
;
immunology
;
prevention & control
;
virology
;
Viral Vaccines
;
administration & dosage
;
genetics
;
immunology
7.Assessing PCV2 antibodies in field pigs vaccinated with different porcine circovirus 2 vaccines using two commercial ELISA systems.
Min Kyoung SHIN ; Seung Hyun YOON ; Myung Hwui KIM ; Young Soo LYOO ; Seung Won SUH ; Han Sang YOO
Journal of Veterinary Science 2015;16(1):25-29
Porcine circovirus type 2 (PCV2) is the primary causative agent for post-weaning, multisystemic, wasting syndrome. Consequently, serologic detection of and vaccination against PCV2 are important for the swine industry. Among several serological tests, the enzyme-linked immunosorbent assay (ELISA) is commonly used to measure anti-PCV2 antibody levels. In the present study, we used two commercial ELISA systems to comparatively evaluate anti-PCV2 antibodies in field pigs treated with three different PCV2 vaccines. Among a total of 517 serum samples, the results of the two ELISAs were fully concordant for 365 positive and 42 negative samples, indicating 78.7% agreement. In addition, the Pearson coefficient (0.636) indicated a moderate correlation between data from the two ELISAs. Results from the farms with pigs vaccinated with the three different PCV2 vaccines demonstrated that most of the vaccinated animals underwent seroconversion. However, the increase and duration of antibody titers varied depending on the vaccine, the presence of maternal antibodies, and the vaccination program. PCV2 serologic status and anti-PCV2 antibody levels of herds from this study could be utilized to determine the best timing for vaccination and assessing vaccination compliance.
Aging
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Animals
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Antibodies, Viral/*blood
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Circovirus/*classification/immunology
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Enzyme-Linked Immunosorbent Assay/methods/*veterinary
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Female
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Porcine Postweaning Multisystemic Wasting Syndrome/blood/immunology/*prevention & control
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Republic of Korea/epidemiology
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Swine
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Swine Diseases/*prevention & control/virology
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Viral Vaccines/*immunology
8.Porcine circovirus type 2 and PCV2-systemic disease--a review.
Jinyan GU ; Gang XING ; Jing LEI ; Fei LIU ; Jiyong ZHOU
Chinese Journal of Biotechnology 2015;31(6):880-891
Porcine circovirus type 2 (PCV2) can cause immunosuppression on herds. PCV2, as an essential pathogen of PCV2-systemic disease (PCV2-SD), has caused considerable economic losses in pig industry worldwide. Here we review and address the evolution, viral protein and immunolesion of PCV2 and preventive techniques of PCV2-SD.
Animals
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Circoviridae Infections
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veterinary
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Circovirus
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genetics
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Phylogeny
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Swine
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Swine Diseases
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virology
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Viral Proteins
;
genetics
9.Cloning and Sequence Analyses of Genome of Swine Hepatitis E Virus(HEV) HN-JY40 Strains Isolated from Henan Province.
Xiaoxia ZHANG ; Qin ZHANG ; Zhenpu LIANG ; Feng XU ; Xinfeng SHAO
Chinese Journal of Virology 2015;31(3):231-238
In the present study, the genomic sequence characteristics of HN-JY40 strains of the hepatitis E virus (HEV) isolated from pigs in Henan Province, China, were analyzed and the evolutionary relationship between HN-JY40 and other sequenced strains examined. The whole genome of HN-JY40 was sequenced and analyzed by reverse transcription-polymerase chain reaction, 3' rapid amplification of cDNA ends (3' RACE) and 5' RACE. Bioinformatic analyses were carried out with Megalign, Expasy, clustal x, and MEGA 4 software. The genome of HN-JY40 was 7 223 bp in size upon removal of polyA sequences. Sizes were 9 bp and 69 bp at 5' and 3' noncoding regions, respectively. The genome of HN-JY40 was predicted to contain three open reading frames (ORFs): ORF1 (5 124 bp) encoding 1 707 amino acids; ORF2 (2 025 bp) encoding 674 amino acids; ORF3 (345 bp) encoding 114 amino acids. Phylogenetic-tree analyses indicated that HN-JY40 is a typical type-IV virus that belongs to a new subgenotype of HEV genotype 4. We sequenced and analyzed the whole genome of HN-JY40. This strategy elicited the genomic characteristics of the HEV isolated from pigs in Henan Province as well as the evolutionary relationships between HN- JY40 and other HEV isolates from pigs. We revealed that the ORF1 of HN-JY40 (153-432 nt) and human HK 104-2004 had high similarity, which offers molecular evidence for uncovering the interspecies transmission of the HEV.
Amino Acid Sequence
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Animals
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Base Sequence
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China
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Cloning, Molecular
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Genome, Viral
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Hepatitis E
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veterinary
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virology
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Hepatitis E virus
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classification
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genetics
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isolation & purification
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Humans
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Molecular Sequence Data
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Open Reading Frames
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Phylogeny
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Sequence Homology, Amino Acid
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Swine
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Swine Diseases
;
virology
10.Construction and identification of a recombinant PRRSV expressing ORF2 of porcine circovirus type 2.
Tingjie ZHANG ; Xing LIU ; Tao SUN ; Xuejiao ZHU ; Baochao FAN ; Juan BAI ; Ping JIANG
Chinese Journal of Virology 2015;31(1):65-73
Porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) are very two important pathogens that have coursed huge economic losses in swine production in worldwide. In this study,a vector pCMV-TJM containing the full-length cDNA clone of PRRSV attenuated strain TJM-F92 was firstly constructed by PCR method. Then a gene sequence containing Afl II/Mlu I e restriction enzyme sites and a transcription regulatory sequence for ORF6 (TRS6) was inserted be- tween ORF7 and 3'UTR, yielding a expression vector pCMV-TJM-TRS. Subsequently, a plasmid pCMV-TJM-Cap was constructed by cloning of PCV2 ORF2 gene into the unique sites Afl II /Mlu I of pCMV- TJM-TRS plasmid DNA. Then three recombinant PRRSV, rTJM, rTJM/TRS and rTJM/Cap, were rescued by transfection of pCMV-TJM, pCMV-TJM-TRS and pCMV-TJM-Cap into Marc-145 cells, respectively,and confirmed by the genome sequence, restriction enzyme digestion, Western Blot and IFA. They all had the molecular markers which was different from the parent virus. The growth characteristics of the rescued viruses were similar to that of parent virus. rTJM/Cap could also express efficiently PCV2 Cap protein in Marc-145 cells. At passage 8, it still had PCV2 ORF2 gene which examined by RT-PCR. It indicated that the full-length cDNA clone of PRRSV attenuated strain TJM-F92 and recombinant PRRSV rTJM/Cap expressing PCV2 Cap protein were successfully constructed. It made an important foundation for studying on the pathogenic mechanisms of PRRSV and PRRSV-PCV2 vaccine in the future.
Animals
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Capsid Proteins
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genetics
;
immunology
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Cell Line
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Circoviridae Infections
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veterinary
;
virology
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Circovirus
;
classification
;
genetics
;
metabolism
;
Gene Expression
;
Open Reading Frames
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Porcine Reproductive and Respiratory Syndrome
;
virology
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Porcine respiratory and reproductive syndrome virus
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genetics
;
metabolism
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Recombination, Genetic
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Swine
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Swine Diseases
;
virology
;
Viral Vaccines
;
genetics
;
immunology

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