Abstract
BackgroundThe US11 protein of herpes simplex virus type 1 (HSV-1) is a small, highly basic phosphoprotein expressed at late times during infection. To date, the function of US11 protein in cell culture and animal models is poorly understood. To further investigate the function of the US11 protein, this study was undertaken to express the US11 protein and raise a polyclonal antibody.ResultsThe US11 gene was cloned into the prokaryotic expression vector pET-32a (+) to express His-tagged US11 protein in Escherichia coli. After purification by nickel affinity chromatography and refolding, the recombinant protein was used to raise the anti-US11 polyclonal antibody. Western blot analysis demonstrated that the US11 protein was specifically recognized by the polyclonal antibody, and immunofluorescent assay also showed that the antibody was able to probe the US11 protein in the cells infected with HSV-1.ConclusionsIn the present study, we obtained a high-level expression of the recombinant US11 protein as well as high titers of rabbit polyclonal antibody specially against US11 protein in HSV-1 infected cells. This special polyclonal antibody provides a good tool for further studying structural and functional characterization of HSV-1 US11 protein.
Highlights
The US11 protein of herpes simplex virus type 1 (HSV-1) is a small, highly basic phosphoprotein expressed at late times during infection
The 152-kb double-stranded Herpes simplex virus type 1 (HSV-1) genome is rapidly translocated to the nucleus where at least 80 viral genes are transcribed by the host cell RNA polymerase II (Pol II) [1]
Expression of the His-tagged US11 protein After induction with 1.0 mM IPTG at 37°C for 4h, E. coli BL21 (DE3) harboring pET-32a-US11 exhibited a high level of expression (Figure 2A, lane 3)
Summary
The US11 protein of herpes simplex virus type 1 (HSV-1) is a small, highly basic phosphoprotein expressed at late times during infection. The 152-kb double-stranded HSV-1 genome is rapidly translocated to the nucleus where at least 80 viral genes are transcribed by the host cell RNA polymerase II (Pol II) [1]. The US11 protein is a 21 kDa, highly basic phosphoprotein [4], and is an RNA-binding protein, post-transcriptional regulator of gene expression [5,6,7]. US11 has been reported as a potent inhibitor of PKR activation through binding to dsRNA [14] or through direct interaction with PKR in the context of viral infection [12] and could interfere with the PKR mediated host cell responses. It is clear that US11 is a multifunctional protein involved in HSV-1 infection
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