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      Replacement of the heterologous 5 untranslated region allows preservation of the fully functional activities of type 2 porcine reproductive and respiratory syndrome virus

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          Abstract

          The 5′ untranslated region (UTR) is believed to be vital for the replication of porcine reproductive and respiratory syndrome virus (PRRSV), yet its functional mechanism remains largely unknown. In this study, to define the cis-acting elements for viral replication and infectivity, The 5′ UTR swapping chimeric clones pTLV8 and pSHSP5 were constructed based on two different genotypes full-length infectious cDNA clone pAPRRS and pSHE backbones. Between them, vTLV8 could be rescued from pTLV8 and had similar virological properties to vAPRRS, including phenotypic characteristic and RNA synthesis level. However, pSHSP5 exhibited no evidence of infectivity. Taken together, the results presented here demonstrate that only the 5′ UTR of type 1 PRRSV did not affect the infectivity and replication of type 2 PRRSV in vitro. The 5′ UTR of type 2 PRRSV could be functionally replaced by its counterpart from type 1.

          Highlights

          ► The 5′ UTR of inter-genotypic swapping mutation have different consequences of viral rescue, RNA synthesis and protein expression. ► Replacement of the type 1 5′ UTR allows preservation of the fully functional activities of type 2 PRRSV, while converse substitution cause chimera lethal. ► The virological properties and RNA synthesis level of vTLV8 are almost indistinguishable from those of the parental virus. ► Type 1 5′ UTR chimeric virus vTLV8 is genetically stable and possesses nucleotides mutations in the nsp9 region.

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          Most cited references43

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          Nidovirales: a new order comprising Coronaviridae and Arteriviridae.

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            The molecular biology of arteriviruses.

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              Lelystad Virus, the Causative Agent of Porcine Epidemic Abortion and Respiratory Syndrome (PEARS), Is Related to LDV and EAV

              The genome of Lelystad virus (LV), the causative agent of porcine epidemic abortion and respiratory syndrome (previously known as mystery swine disease), was shown to be a polyadenylated RNA molecule. The nucleotide sequence of the LV genome was determined from a set of overlapping cDNA clones. A consecutive sequence of 15,088 nucleotides was obtained. Eight open reading frames (ORFs) that might encode virus-specific proteins were identified. ORF1a and ORF1b are predicted to encode the vital RNA polymerase because the amino acid sequence contains sequence elements that are conserved in RNA polymerases of the torovirus Berne virus (BEV), equine arteritis virus (EAV), lactate dehydrogenase-elevating virus (LDV), the coronaviruses, and other positive-strand RNA viruses. A heptanucleotide slippery sequence (UUUAAAC) and a putative pseudoknot structure, which are both required for efficient ribosomal frameshifting during translation of the RNA polymerase ORF 1b of BEV, EAV, and the coronaviruses, were identified in the overlapping region of ORF1a and ORF1b of LV. ORFs 2 to 6 probably encode viral membrane-associated proteins, whereas ORF7 is predicted to encode the nucleocapsid protein. Comparison of the amino acid sequences of the ORFs identified in the genome of LV, LDV, and EAV indicated that LV and LDV are more closely related than LV and EAV. A 3′ nested set of six subgenomic RNAs was detected in LV-infected cells. These subgenomic RNAs contain a common leader sequence that is derived from the 5′ end of the genomic RNA and that is joined to the 3′ terminal body sequence. Our results indicate that LV is closely related evolutionarily to LDV and EAV, both members of a recently proposed family of positive-strand RNA viruses, the Arteriviridae.
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                Author and article information

                Contributors
                Journal
                Virology
                Virology
                Virology
                Elsevier Inc.
                0042-6822
                1096-0341
                1 March 2013
                25 April 2013
                1 March 2013
                : 439
                : 1
                : 1-12
                Affiliations
                [a ]Department of Swine Infectious Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, PR China
                [b ]College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province 210095, PR China
                Author notes
                [* ]Corresponding author. Tel.: +86 21 3429 3156. gztong@ 123456shvri.ac.cn shishanyuan@ 123456hotmail.com
                Article
                S0042-6822(12)00641-1
                10.1016/j.virol.2012.12.013
                7111940
                23453581
                c86c6348-72fd-4bdd-95be-9f902ac0757f
                Copyright © 2013 Elsevier Inc. All rights reserved.

                Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

                History
                : 30 September 2012
                : 20 December 2012
                : 25 December 2012
                Categories
                Article

                Microbiology & Virology
                prrsv,5′ utr,chimeric clone,stem–loop structure
                Microbiology & Virology
                prrsv, 5′ utr, chimeric clone, stem–loop structure

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