title: Preparation and characterization of an antiserum against truncated UL54 protein of pseudorabies virus.
abstract: Pseudorabies virus (PRV) early protein UL54 is a homolog of herpes simplex virus 1 immediate-early protein ICP27, which is a multifunctional protein essential for the virus replication. However, the precise role of the PRV UL54 protein in the virus life cycle is still poorly understood. To shed more light on this problem, we considered it essential to have available an antiserum specifically detecting this protein. Since it was known that a full-length UL54 protein is a too big molecule for efficient expression in prokaryotic systems, it was truncated from 1 to 66 N-terminal amino acids, fused to EYFP-His tag and expressed in Escherichia coli through an appropriate expression vector. The truncated protein was purified by Ni-NTA affinity chromatography and used for raising an antiserum in rabbits. Western blot analysis showed that this antiserum specifically recognized the purified truncated as well as full-length UL54 protein in PRV-infected cells. Immunofluorescence assay confirmed the latter finding and also demonstrated localization of this protein first in nucleoli and later in whole nuclei of PRV-infected cells. These results indicate that the prepared antiserum could serve as a valuable tool in further studies of PRV UL54 protein function.
authors.affiliation: Department of Pathogenic Biology and Immunology, Guangzhou Medical University, People's Republic of China.
authors.forename: M L
authors.initials: ML
authors.lastname: Li
authors.forename: Z
authors.initials: Z
authors.lastname: Li
authors.forename: W T
authors.initials: WT
authors.lastname: Li
authors.forename: B Y
authors.initials: BY
authors.lastname: Wang
authors.forename: C Q
authors.initials: CQ
authors.lastname: Ma
authors.forename: J H
authors.initials: JH
authors.lastname: Chen
authors.forename: M S
authors.initials: MS
authors.lastname: Cai
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D002460:Cell Line; D011558:Herpesvirus 1, Suid / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D006801:Humans; D007106:Immune Sera / Q000276:immunology*; D011557:Pseudorabies / Q000175:diagnosis / Q000276:immunology / Q000821:virology; D011817:Rabbits; D017384:Sequence Deletion; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D000914:Antibodies, Viral; D007106:Immune Sera; D014764:Viral Proteins