title: Identification and characterization of the duck enteritis virus (DEV) US2 gene.
abstract: The US2 protein has been reported to contribute to duck enteritis virus (DEV) infection; however, its kinetics and localization during infection, and whether it is a component of virion, have not been previously reported. To elucidate the function of DEV US2, US2 was amplified by polymerase chain reaction (PCR) and inserted into pET-32a(+); this was expressed, the recombinant US2 protein was purified, and a polyclonal antibody generated. In addition, the kinetics and localization of the US2 gene and protein were determined by quantitative real-time fluorescent PCR, ganciclovir (GCV), and cycloheximide (CHX) treatment, western-blot, and indirect immunofluorescence assay. The packaging of US2 into DEV virions was revealed by a protease protection assay. US2 was found to be transcribed 24 h post-infection (pi) and peaked at 72 h pi; the US2 protein was detected 48 h pi, except in the presence of GCV or CHX. US2 was packed into virions and also localized to the plasma membrane and cytoplasm in infected cells. The results showed that the DEV US2 is a late gene, and that its encoding protein could be a tegument component localized mainly in the cytoplasm. This study provides useful data for the further analysis of DEV US2, including an explanation for the genetic conservation among alphaherpesviruses.
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu, Sichuan, China.
authors.forename: J
authors.initials: J
authors.lastname: Gao
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu, Sichuan, China chenganchun@vip.163.com.
authors.forename: A C
authors.initials: AC
authors.lastname: Cheng
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu, Sichuan, China.
authors.forename: M S
authors.initials: MS
authors.lastname: Wang
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu, Sichuan, China.
authors.forename: R Y
authors.initials: RY
authors.lastname: Jia
authors.affiliation: Avian Disease Research Center, College of Veterinary Medicine of Sichuan Agricultural University, Wenjiang, Chengdu, Sichuan, China.
authors.forename: D K
authors.initials: DK
authors.lastname: Zhu
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu, Sichuan, China.
authors.forename: S
authors.initials: S
authors.lastname: Chen
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu, Sichuan, China.
authors.forename: M F
authors.initials: MF
authors.lastname: Liu
authors.affiliation: Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Wenjiang, Chengdu, Sichuan, China.
authors.forename: F
authors.initials: F
authors.lastname: Liu
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu, Sichuan, China.
authors.forename: Q
authors.initials: Q
authors.lastname: Yang
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu, Sichuan, China.
authors.forename: K F
authors.initials: KF
authors.lastname: Sun
authors.affiliation: Avian Disease Research Center, College of Veterinary Medicine of Sichuan Agricultural University, Wenjiang, Chengdu, Sichuan, China.
authors.forename: X Y
authors.initials: XY
authors.lastname: Chen
mesh_terms: D000818:Animals; D002460:Cell Line; D004372:Ducks; D005347:Fibroblasts; D015870:Gene Expression; D025922:Mardivirus / Q000187:drug effects / Q000235:genetics*; D021381:Protein Transport; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D014771:Virion
chemical_list: D011994:Recombinant Proteins; D014759:Viral Envelope Proteins