Molecular Characterization of UL50 (dUTPase) Gene of Bovine Herpes Virus 1
Bovine herpes virus -1 (BoHV-1) infection leads to upper respiratory tract infection, conjunctivitis and genital disorders in cattle. To control BoHV-1, it is important to understand the role of viral proteins in viral infection. BoHV-1 has several gene products to help in viral replication in infec...
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| Format: | Article |
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MDPI AG
2023-08-01
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| Series: | Animals |
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| Online Access: | https://www.mdpi.com/2076-2615/13/16/2607 |
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| author | Farzana Shahin Sohail Raza Xi Chen Changmin Hu Yingyu Chen Huanchun Chen Aizhen Guo |
| author_facet | Farzana Shahin Sohail Raza Xi Chen Changmin Hu Yingyu Chen Huanchun Chen Aizhen Guo |
| author_sort | Farzana Shahin |
| collection | DOAJ |
| description | Bovine herpes virus -1 (BoHV-1) infection leads to upper respiratory tract infection, conjunctivitis and genital disorders in cattle. To control BoHV-1, it is important to understand the role of viral proteins in viral infection. BoHV-1 has several gene products to help in viral replication in infected cell. One such gene is deoxyuridine triphosphate nucleotidohydrolase (dUTPase) also known as UL50. In this study, we analyzed the amino acid sequence of UL50 (dUTPase) using bioinformatics tools and found that it was highly conserved among herpesvirus family. Then, it was cloned and expressed in <i>Escherichia coli</i> Rosetta (DE3), induced by isopropy1-b-D-thiogalactopyranoside (IPTG) and the recombinant UL50 protein was purified to immunize rabbits for the preparation of polyclonal antiserum. The results indicated that the UL50 gene of BoHV-1 was composed of 978 nucleotides, which encoded 323 amino acids. Western blot analysis revealed that polyclonal sera against UL50 reacted with a band of 34 kDa. Furthermore, immunofluorescence assay showed that UL50 localized in the cytoplasmic area. Taken together, UL50 was successfully cloned, expressed and detected in BoHV-1-infected cells and was localized in the cytoplasm to help in the replication of BoHV-1 in infected cells. |
| first_indexed | 2024-03-11T00:11:09Z |
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| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-11T00:11:09Z |
| publishDate | 2023-08-01 |
| publisher | MDPI AG |
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| series | Animals |
| spelling | doaj.art-07db5e41f602411191de60ff02def2bc2023-11-18T23:57:37ZengMDPI AGAnimals2076-26152023-08-011316260710.3390/ani13162607Molecular Characterization of UL50 (dUTPase) Gene of Bovine Herpes Virus 1Farzana Shahin0Sohail Raza1Xi Chen2Changmin Hu3Yingyu Chen4Huanchun Chen5Aizhen Guo6The State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, ChinaThe State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, ChinaCollege of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, ChinaCollege of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, ChinaCollege of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, ChinaThe State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, ChinaThe State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, ChinaBovine herpes virus -1 (BoHV-1) infection leads to upper respiratory tract infection, conjunctivitis and genital disorders in cattle. To control BoHV-1, it is important to understand the role of viral proteins in viral infection. BoHV-1 has several gene products to help in viral replication in infected cell. One such gene is deoxyuridine triphosphate nucleotidohydrolase (dUTPase) also known as UL50. In this study, we analyzed the amino acid sequence of UL50 (dUTPase) using bioinformatics tools and found that it was highly conserved among herpesvirus family. Then, it was cloned and expressed in <i>Escherichia coli</i> Rosetta (DE3), induced by isopropy1-b-D-thiogalactopyranoside (IPTG) and the recombinant UL50 protein was purified to immunize rabbits for the preparation of polyclonal antiserum. The results indicated that the UL50 gene of BoHV-1 was composed of 978 nucleotides, which encoded 323 amino acids. Western blot analysis revealed that polyclonal sera against UL50 reacted with a band of 34 kDa. Furthermore, immunofluorescence assay showed that UL50 localized in the cytoplasmic area. Taken together, UL50 was successfully cloned, expressed and detected in BoHV-1-infected cells and was localized in the cytoplasm to help in the replication of BoHV-1 in infected cells.https://www.mdpi.com/2076-2615/13/16/2607BoHV-1dUTPasepolyclonal antiserumclonedlocalizedUL50 |
| spellingShingle | Farzana Shahin Sohail Raza Xi Chen Changmin Hu Yingyu Chen Huanchun Chen Aizhen Guo Molecular Characterization of UL50 (dUTPase) Gene of Bovine Herpes Virus 1 Animals BoHV-1 dUTPase polyclonal antiserum cloned localized UL50 |
| title | Molecular Characterization of UL50 (dUTPase) Gene of Bovine Herpes Virus 1 |
| title_full | Molecular Characterization of UL50 (dUTPase) Gene of Bovine Herpes Virus 1 |
| title_fullStr | Molecular Characterization of UL50 (dUTPase) Gene of Bovine Herpes Virus 1 |
| title_full_unstemmed | Molecular Characterization of UL50 (dUTPase) Gene of Bovine Herpes Virus 1 |
| title_short | Molecular Characterization of UL50 (dUTPase) Gene of Bovine Herpes Virus 1 |
| title_sort | molecular characterization of ul50 dutpase gene of bovine herpes virus 1 |
| topic | BoHV-1 dUTPase polyclonal antiserum cloned localized UL50 |
| url | https://www.mdpi.com/2076-2615/13/16/2607 |
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