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Aims: Penaeus merguiensis hepandensovirus (PmeHDV) (GenBank No. DQ458781) is a shrimp hepatopancreatic parvovirus (HPV), belonging to subfamily Densovirinae. Transportation of Densovirinae into and out of nucleus is allowed by the binding of nuclear location signals (NLSs) to importins (Imp). PmeHDV has putative NLSs that need to be experimentally tested. The aims of this study is to determine if the three putative NLSs of PmeHDV are functioning by transfecting NLS-inserted-plasmid DNAs into Vero cell lines using a transfection reagent.
Place and Duration of the Study: Data for this study was collected from the Veterinary and Biomedical Sciences Laboratories at James Cook University (JCU) during the duration from May 2015 to December 2016. Methodology: Each plasmid has been synthetically inserted with each sequence of the putative NLSs and a
fluorescent protein. The presence of the NLS in the cell nucleus and cytoplasm was screened. The overlay of visualization of transfected plasmids is presented.
Results: It appears the NLSs are not functioning well as that the proteins are blocked at the nuclear membrane, probably linked to importin beta-1 and not frequently entering the nucleus. Our study demonstrated small noticeable differences in the outer nuclei within transfected-Vero cells with the experimental NLSs genes.
Conclusion: In conclusion, our fluorescent study was not sufficiently sensitive to be confident of the detection in NLS-transfected cells under different filters. The study of crustacean virus-host interactions using proxy cell cultures as models remains a major challenge.