title: Expression and characterization of UL16 gene from duck enteritis virus.
abstract: BACKGROUND
  Previous studies have indicated that the UL16 protein and its homologs from herpesvirus were conserved and played similar roles in viral DNA packaging, virion assembly, budding, and egress. However, there was no report on the UL16 gene product of duck enteritis virus (DEV). In this study, we analyzed the amino acid sequence of UL16 using bioinformatics tools and expressed in Escherichia coli Rosetta (DE3) induced by isopropy1-β-D-thiogalactopyranoside (IPTG). The recombinant protein was produced, purified using a Ni-NTA column and used to generate the polyclonal antibody against UL16. The intracellular distribution of the DEV UL16 product was carried out using indirect immunofluorescence assay.
  
  
  RESULTS
  In our study, UL16 gene of DEV was composed of 1089 nucleotides, which encoded 362 amino acids. Multiple sequence alignment suggested that the UL16 gene was highly conserved in herpesvirus family. The UL16 gene was cloned into a pET prokaryotic expression vector and transformed into Escherichia coli Rossetta (DE3) induced by IPTG. A 60kDa fusion protein band corresponding to the predicted size was produced on the SDS-PAGE, purified using a Ni-NTA column. Anti-UL16 polyclonal sera was prepared by immunizing rabbits, and reacted with a band in the IPTG induced cell lysates with an apparent molecular mass of 60 kDa. In vivo expression of the UL16 protein in DEV infected duck embryo fibroblast cells (DEFs) was localized mostly around perinuclear cytoplasmic area and in cytosol using indirect immunofluorescence assay.
  
  
  CONCLUSIONS
  The UL16 gene of DEV was successfully cloned, expressed and detected in DEV infected DEFs for the first time. The UL16 protein localized mostly around perinuclear cytoplasmic area and in cytosol in DEV infected DEFs. DEV UL16 shared high similarity with UL16 family members, indicating that DEV UL16 many has similar function with its homologs. All these results may provide some insight for further research about full characterizations and functions of the DEV UL16.
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu City, Sichuan 611130, P.R. China.
authors.forename: Qin
authors.initials: Q
authors.lastname: He
authors.forename: Qiao
authors.initials: Q
authors.lastname: Yang
authors.forename: Anchun
authors.initials: A
authors.lastname: Cheng
authors.forename: Mingshu
authors.initials: M
authors.lastname: Wang
authors.forename: Jun
authors.initials: J
authors.lastname: Xiang
authors.forename: Dekang
authors.initials: D
authors.lastname: Zhu
authors.forename: Renyong
authors.initials: R
authors.lastname: Jia
authors.forename: Qihui
authors.initials: Q
authors.lastname: Luo
authors.forename: Zhengli
authors.initials: Z
authors.lastname: Chen
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhou
authors.forename: Xiaoyue
authors.initials: X
authors.lastname: Chen
mesh_terms: D018138:Alphaherpesvirinae / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000378:metabolism; D001715:Bird Diseases / Q000473:pathology / Q000821:virology*; D003001:Cloning, Molecular; D017124:Conserved Sequence / Q000235:genetics; D004372:Ducks / Q000821:virology*; D004591:Electrophoresis, Polyacrylamide Gel; D004751:Enteritis / Q000473:pathology / Q000821:virology*; D004926:Escherichia coli; D005347:Fibroblasts / Q000166:cytology / Q000821:virology; D015870:Gene Expression; D006566:Herpesviridae Infections / Q000473:pathology / Q000821:virology*; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D010957:Plasmids; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D016415:Sequence Alignment; D014169:Transformation, Bacterial; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D000906:Antibodies; D011994:Recombinant Proteins; D014764:Viral Proteins