GENOME ANNOUNCEMENT
Complete Genome Sequence of a Porcine Orthoreovirus from Southern China Yimin Dai,a Qingfeng Zhou,c Chengwen Zhang,c Yanhua Song,c Xiaoyan Tian,c Xiangbin Zhang,c Chunyi Xue,a Shun Xu,a Yingzuo Bi,b and Yongchang Caoa State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, People’s Republic of Chinaa; College of Animal Science, South China Agricultural University, Guangzhou, People’s Republic of Chinab; and Guangdong Wen’s Group Academy, Guangdong Wen’s Foodstuffs Group Co., Ltd., Xinxing, Guangdong, People’s Republic of Chinac
Porcine orthoreoviruses belong to the family Reoviridae and cause mainly mild enteritis in piglets. We present here the complete genome sequence of a novel porcine orthoreovirus strain (GD-1) isolated from a piglet in southern China. Our data will facilitate future investigations of the molecular characteristics and epidemiology of porcine orthoreoviruses.
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orcine orthoreoviruses belong to mammalian orthoreoviruses and are members of the Reoviridae family (1), which is a family of nonenveloped, double-stranded RNA viruses. The mammalian orthoreovirus genome is composed of 10 segments, 3 large (L1 to L3), 3 medium (M1 to M3), and 4 small (S1 to S4) (6, 7, 9). The virus genome encodes eight structural proteins (1 to 3, 1 to 3, 1, and 2) and four nonstructural proteins (NS, NSC, NS, and 1s) (6). Mammalian orthoreoviruses can be divided into three serotypes classically through neutralization and hemagglutination inhibition tests (3, 5, 8), whose representative prototypes are type 1 Lang (T1L), type 2 Jones (T2J), and type 3 Dearing (T3D), respectively (2, 4–6). Here we report the complete genome sequence of porcine orthoreovirus strain GD-1, which was isolated from a piglet with diarrhea in Guangdong Province in southern China. Virus RNA was extracted from Vero cells infected with GD-1 and then used for full-length amplification of cDNAs. Ten pairs of primers based on the terminal conserved sequence of the orthoreovirus genome were used to amplify the GD-1 genome. The PCR products were gel purified and cloned into the simple vector pMD-18T (TaKaRa). Thirteen primers, in addition to universal primers M13F and M13R, were used to determine the whole GD-1 genome sequence with an ABI Prism 3730 sequencer (Applied Biosystems), and then the sequences were assembled and manually edited to obtain the final full-length genome sequence. The complete genome of GD-1 was 23,539 bp in length, including segments L1 to L3, M1 to M3, and S1 to S4 (3,901 bp, 3,915 bp, 3,854 bp, 2,241 bp, 2,203 bp, 2,304 bp, 1,416 bp, 1,331 bp, 1,198 bp, and 1,196 bp, respectively), encoding proteins 1 (1,275 amino acids [aa]), 2 (1,289 aa), 3 (1,267 aa), NS (721 aa), NSC (681 aa), 1 (708 aa), 2 (736 aa), 1 (455 aa), 1S(119 to 125 aa), 2 (418 aa), NS (375 aa), and 3 (365 aa). Based on homology analysis of the deduced S1 amino acid sequences of GD-1 and reference strains T1L, T2J, and T3D, strain GD-1 was identified as reovirus type 3. Among the 10 segments of GD-1, L1, L2, and L3 show the highest levels of identity (96%, 97%, and 98%, respectively) with strain T3D; M1 and S3 of GD-1 share up to 95% and 93% identity with their T1L counterparts.
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Furthermore, the sequence of S4 of GD-1 is significantly different among T1L, T2J, and T3D, which share just 82% identity with T2J. Nucleotide sequence accession numbers. The complete genome sequence of strain GD-1 has been deposited in GenBank under accession numbers JX486057 to JX486066. ACKNOWLEDGMENTS This work was supported by grants from the State Key Laboratory of Biocontrol at Sun Yat-sen University and WENS Foundation grants from Guangdong Wen’s Foodstuffs Group Co., Ltd. We thank to George D. Liu for critical review and revision of the manuscript.
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Received 23 August 2012 Accepted 23 August 2012 Address correspondence to Yongchang Cao,
[email protected]. Copyright © 2012, American Society for Microbiology. All Rights Reserved. doi:10.1128/JVI.02254-12
November 2012 Volume 86 Number 22
