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      Evaluation in Swine of a Recombinant African Swine Fever Virus Lacking the MGF-360-1L Gene

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          Abstract

          The African swine fever (ASF) pandemic is currently affecting pigs throughout Eurasia, resulting in significant swine production losses. The causative agent, ASF virus (ASFV), is a large, structurally complex virus with a genome encoding more than 160 genes. The function of most of those genes remains unknown. Here, we presented the previously uncharacterized ASFV gene MGF360-1L, the first gene in the genome. The kinetic studies of virus RNA transcription demonstrated that the MGF360-1L gene was transcribed as a late virus protein. The essentiality of MGF360-1L to virus replication was evaluated by developing a recombinant ASFV lacking the gene (ASFV-G-ΔMGF360-1L). In primary swine macrophage cell cultures, ASFV-G-ΔMGF360-1L showed similar replication kinetics as the parental highly virulent field isolate Georgia2007 (ASFV-G). Domestic pigs experimentally infected with ASFV-G-ΔMGF360-1L presented with a clinical disease indistinguishable from that caused by ASFV-G, demonstrating that MGF360-1L was not involved in virulence in swine, the natural host of ASFV.

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          Most cited references36

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          Purification of RNA using TRIzol (TRI reagent).

          TRIzol solubilization and extraction is a relatively recently developed general method for deproteinizing RNA. This method is particularly advantageous in situations where cells or tissues are enriched for endogenous RNases or when separation of cytoplasmic RNA from nuclear RNA is impractical. TRIzol (or TRI Reagent) is a monophasic solution of phenol and guanidinium isothiocyanate that simultaneously solubilizes biological material and denatures protein. After solubilization, the addition of chloroform causes phase separation (much like extraction with phenol:chloroform:isoamyl alcohol), where protein is extracted to the organic phase, DNA resolves at the interface, and RNA remains in the aqueous phase. Therefore, RNA, DNA, and protein can be purified from a single sample (hence, the name TRIzol). TRIzol extraction is also an effective method for isolating small RNAs, such as microRNAs, piwi-associated RNAs, or endogeneous, small interfering RNAs. However, TRIzol is expensive and RNA pellets can be difficult to resuspend. Thus, the use of TRIzol is not recommend when regular phenol extraction is practical.
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            A Mouse Model of Zika Virus Pathogenesis.

            The ongoing Zika virus (ZIKV) epidemic and unexpected clinical outcomes, including Guillain-Barré syndrome and birth defects, has brought an urgent need for animal models. We evaluated infection and pathogenesis with contemporary and historical ZIKV strains in immunocompetent mice and mice lacking components of the antiviral response. Four- to six-week-old Irf3(-/-)Irf5(-/-)Irf7(-/-) triple knockout mice, which produce little interferon α/β, and mice lacking the interferon receptor (Ifnar1(-/-)) developed neurological disease and succumbed to ZIKV infection, whereas single Irf3(-/-), Irf5(-/-), and Mavs(-/-) knockout mice exhibited no overt illness. Ifnar1(-/-) mice sustained high viral loads in the brain and spinal cord, consistent with evidence that ZIKV causes neurodevelopmental defects in human fetuses. The testes of Ifnar1(-/-) mice had the highest viral loads, which is relevant to sexual transmission of ZIKV. This model of ZIKV pathogenesis will be valuable for evaluating vaccines and therapeutics as well as understanding disease pathogenesis.
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              A live-attenuated Zika virus vaccine candidate induces sterilizing immunity in mouse models

              Pei-Yong Shi and colleagues report that a deletion mutant of Zika virus is safe and effective as a live-attenuated vaccine in mice and induces sterilizing immunity. Their results encourage further testing of the candidate vaccine for possible future use in humans.
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                Author and article information

                Journal
                Viruses
                Viruses
                viruses
                Viruses
                MDPI
                1999-4915
                20 October 2020
                October 2020
                : 12
                : 10
                : 1193
                Affiliations
                [1 ]Plum Island Animal Disease Center, ARS, USDA, Greenport, NY 11944, USA; Elizabeth.Ramirez@ 123456usda.gov (E.R.-M.); Elizabeth.Vuono@ 123456usda.gov (E.A.V.); Ayushi.Rai@ 123456usda.gov (A.R.); Sarah.Pruitt@ 123456usda.gov (S.P.); Ediane.Silva@ 123456usda.gov (E.S.); Lauro.Velazquez@ 123456usda.gov (L.V.-S.); James.Zhu@ 123456usda.gov (J.Z.)
                [2 ]Department of Pathobiology and Veterinary Science, University of Connecticut, Storrs, CT 06269, USA
                [3 ]Department of Pathobiology and Population Medicine, Mississippi State University, P.O. Box 6100, Mississippi, MS 39762, USA
                [4 ]Oak Ridge Institute for Science and Education (ORISE), Oak Ridge, TN 37830, USA
                [5 ]Department of Anatomy and Physiology, Kansas State University, Manhattan, KS 66506, USA
                Author notes
                [* ]Correspondence: douglas.gladue@ 123456usda.gov (D.P.G.); manuel.borca@ 123456usda.gov (M.V.B.); Tel.: +1-631-323-3035 (D.P.G.); +1-631-323-3019 (M.V.B.)
                [†]

                These authors contributed equally to this work.

                Author information
                https://orcid.org/0000-0002-7894-0233
                Article
                viruses-12-01193
                10.3390/v12101193
                7589680
                33092258
                46f88f6f-e4a6-457f-ae59-73df84f0f9a4
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 14 September 2020
                : 16 October 2020
                Categories
                Communication

                Microbiology & Virology
                asfv,asf,african swine fever virus,mgf360-1l
                Microbiology & Virology
                asfv, asf, african swine fever virus, mgf360-1l

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