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Identification of dispensable nucleotide sequence in 3' untranslated region of porcine reproductive and respiratory syndrome virus.

ABSTRACT: The 3' untranslated region (UTR) of porcine arterivirus genome plays a pivotal role for virus replication, yet the properties of 3' UTR remain largely undefined. We conducted site-directed mutagenesis to the 3' UTR of the type II porcine reproductive and respiratory syndrome virus (PRRSV). Serial deletions of the 3' UTR showed that at least 40 nucleotides immediately following the ORF7 stop codon were dispensable for the viability of PRRSV in cultured cells. We then constructed a chimeric PRRSV cDNA clone using type II PRRSV as the backbone containing the 3' UTR from the type I PRRSV. The chimeric virus was viable and shared similar properties with the parental virus. Our results provided the first description of the 40nt dispensable region in type I PRRSV 3' UTR, and further predicted structure demonstrated that the high-order structure of 3' UTR might play significant roles in its function.

SUBMITTER: Sun Z

PROVIDER: S-EPMC7114379 | biostudies-literature | 2010 Dec

REPOSITORIES: biostudies-literature

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Identification of dispensable nucleotide sequence in 3' untranslated region of porcine reproductive and respiratory syndrome virus.

Sun Zhi Z Liu Changlong C Tan Feifei F Gao Fei F Liu Ping P Qin Aijian A Yuan Shishan S

Virus research 20100915 1-2

The 3' untranslated region (UTR) of porcine arterivirus genome plays a pivotal role for virus replication, yet the properties of 3' UTR remain largely undefined. We conducted site-directed mutagenesis to the 3' UTR of the type II porcine reproductive and respiratory syndrome virus (PRRSV). Serial deletions of the 3' UTR showed that at least 40 nucleotides immediately following the ORF7 stop codon were dispensable for the viability of PRRSV in cultured cells. We then constructed a chimeric PRRSV ...[more]

PMID: 20833212

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Project description:BackgroundPorcine reproductive and respiratory syndrome virus (PRRSV) is a causative agent of Porcine Reproductive and Respiratory Syndrome (PRRS). In this study, the complete nucleotide sequences of the selected two Thai PRRSV isolates, EU (01CB1) and US (01NP1) genotypes were determined since both isolates are the Thai prototypes.Results01CB1 and 01NP1 contain 14,943 and 15,412 nucleotides, respectively. The viruses compose 2 untranslated regions (5' UTR and 3' UTR) and 8 open reading frames (ORFs) designated as ORF1a, ORF1b and ORF2-7. Phylogenetic analysis of full length of the viruses also showed that the 01CB1 and 01NP1 were grouped into the EU and US genotype, respectively. In order to determine the genetic variation and genetic relatedness among PRRSV isolates, the complete nucleotide sequences of PRRSV isolated in Thailand, 01CB1 and 01NP1 were compared with those of 2 EU strains (Lelystad, and EuroPRRSV), 6 US strains (MLV, VR2332, PA8, 16244B, SP and HUN4). Our results showed that the 01CB1 genome shares approximately 99.2% (Lelystad) and 95.2% (EuroPRRSV) nucleotide identity with EU field strains. While, the 01NP1 genome has 99.9% nucleotide identity with a live vaccine strain (MLV) and 99.5% and 98.5% nucleotide identity with 2 other US isolates, VR2332 and 16244B, respectively. In addition, ORF5 nucleotide sequences of 9 PRRS viruses recovered in Thailand during 2002-2008 were also included in this study. Phylogenetic analysis of ORF5 showed high similarity among EU and US genotypes of the recent Thai PRRS viruses (2007-2008 viruses) with 01CB1 and 01NP1.ConclusionOverall, the results suggested that the Thai EU isolate (01CB1) may evolve from the EU prototype, Lelystad virus, whereas the Thai US isolate (01NP1) may originate and evolve from the vaccine virus or its derivatives. Interestingly, the US-MLV vaccine was not available in the Thai market in 2001. The Vaccine-like virus might have persisted in the imported pigs or semen and later spread in the Thai swine industry. This report is the first report of complete nucleotide sequences of the Thai PRRS viruses both EU and US genotypes.

Dataset Information

Identification of dispensable nucleotide sequence in 3' untranslated region of porcine reproductive and respiratory syndrome virus.

Publications

Identification of dispensable nucleotide sequence in 3' untranslated region of porcine reproductive and respiratory syndrome virus.

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