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標題: Characterization and expression of the pseudorabies virus early gene UL54
作者: Huang, C.J.;Wu, C.Y.
關鍵字: pseudorabies virus;early gene;immediate-early protein;RNA binding;herpes-simplex-virus;regulatory protein icp27;immediate-early protein;messenger-rna;infected-cells;type-1;transcription;regions;herpesvirus-1;localization
日期: 2004
Issue Date: 2012-12-14 10:28:19 (UTC+8)
關連: Journal of Virological Methods, Volume 119, Issue 2, Page(s) 129-136.
摘要: Pseudorabies virus (PRV) is an alphaherpesvirus, and its gene organization and regulation are similar to the well-characterized human simplex virus (HSV). Sequence analysis of the complete coding region of PRV UL54 gene revealed that the UL54 gene consisted of 1092 nucleotides encoding a protein of 363 amino acids and the gene showed homology to HSV immediate-early protein ICP27. Detection of the UL54 transcript in infected cells by reverse transcription-polymerase chain reaction (RT-PCR) demonstrated that the UL54 gene belonged to the early kinetic class based on sensitivity to cycloheximide, and insensitivity to phosphonoacetic acid (PAA). To study the structure and function of UL54 protein, this gene was subcloned on Escherichia coli expression vector pET28b for overexpression, and the expressed product was applied to generate specific antibody against UL54 protein. The specificity of the mouse immuneserum was confirmed by its ability to react with a 40 kDa viral protein present in the PRV infected cells in Western immunblotting assay, detected as early as 4 h after infection. In addition, immunoperoxidasing staining of PRV infected cells undertaken with this antibody demonstrated mainly nuclear staining pattern. Further-more, the RNA binding potential of UL54 protein was demonstrated by its binding activity to poly(G) RNA homopolymer in Northwestern blotting assay. (C) 2004 Elsevier B.V. All rights reserved.
Relation: Journal of Virological Methods
Appears in Collections:[依資料類型分類] 期刊論文
[依教師分類] 黃千衿

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