Abstract
BackgroundMost of the previous research work had focused on the epidemiology and prevention of duck enteritis virus (DEV). Whilst with the development of protocols in molecular biology, nowadays more and more information about the genes of DEV was reported. But little information about DEV UL53 gene and glycoprotein K(gK) was known except our reported data.ResultsIn our paper, the fluorescent quantitative real-time PCR(FQ-RT-PCR) assay and nucleic acid inhibition test were used to study the transcription characteristic of the DEV UL53 gene. Except detecting the mRNA of DEV UL53 gene, the product gK encoded by UL53 gene was detected through the expression kinetics of UL53 gene by the purified rabbit anti-UL53 protein polyclonal antibodies. Western-blotting and indirect immunofluorescence assays were used to detect gK. From the results of these experiments, the UL53 gene and gK were respectively identified as a late gene and a really late protein. On the other hand, the indirect immunofluorescence assay provided another information that the intracellular localization of DEV gK was mainly distributed in cytoplasm.ConclusionsBy way of conclusions, we conceded that DEV UL53 gene is a really late gene, which is coincident with properties of UL53 homologs from other herpesvirus, such as ILTV(Infectious Laryngotracheitis virus) and HSV-1(Herpes simplex virus type 1). The properties of intracellular localization about gK protein provided a foundation for further functional analysis and further studies will be focused on constructing of the UL53 gene DEV mutant.
Highlights
Most of the previous research work had focused on the epidemiology and prevention of duck enteritis virus (DEV)
Fluorescent quantitative real-time PCR (FQ-RT-PCR) detect the UL53 gene transcript during DEV replication Detect the specificity of the primers and the integrality or purity of the total RNA of each sample The primers P1,P2 for amplifying 164 bp of UL53 gene and primers P3,P4 for amplifying 178 bp of b-actin gene were detected the specificity by traditional PCR
The PCR products were fractionated on 1.5% agarose gel electrophoresis and stained with golden view
Summary
Most of the previous research work had focused on the epidemiology and prevention of duck enteritis virus (DEV). Whilst with the development of protocols in molecular biology, nowadays more and more information about the genes of DEV was reported. With the development of protocols in molecular biology, nowadays more and more information about the genes of DEV was reported, such as UL5 [7], gC [8,9,10], UL24 [11,12,13], UL31 [14,15], UL35 [16,17], UL46 [18], UL38 [19], gE [20], UL51 [21], TK gene [22] and so on. The research will provide useful data for DEV UL53 gene’s properties or gK functional analysis, and will be useful for further understanding the localization properties of alphaherpesvirus UL53 homologs
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