Molecular Characterization of UL50 (dUTPase) Gene of Bovine Herpes Virus 1

SIMPLE SUMMARY: We conducted this study to understand how a virus called bovine herpes virus-1 (BoHV-1) affects cattle. We focused on a specific gene called UL50, which produces a protein called dUTPase. This protein is important for the virus to replicate in infected cells. We analyzed the UL50 gene and found that it was similar across different herpesviruses. We then cloned and expressed the UL50 gene in a bacterium, purified the resulting protein, and used it to create a special antiserum. Through experiments, we discovered that the UL50 protein was present in infected cells and located in the cytoplasm. These findings help us understand how BoHV-1 causes disease in cattle and may aid in developing control measures. ABSTRACT: 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 Escherichia coli 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.