Complete Genomic Sequence of Bluetongue Virus Serotype 16 from China

Abstract

We report here the complete genomic sequence of the Chinese bluetongue virus serotype 16 (BTV16) strain BN96/16. This work is the first to document the complete genomic sequence (segments 1 to 10) of a BTV16 strain. The sequence information provided herein will help determine the geographic origin of BTV16 and define the phylogenetic relationship of BTV16 to other BTV strains.Bluetongue virus (BTV) is the "type" species of the genus Orbivirus, within the family Reoviridae (7). It is the etiological agent of bluetongue disease, which is listed as a "notifiable disease" by the Office International des Epizooties (OIE). The genome of BTV consists of 10 linear double-stranded RNA (dsRNA) segments encoding seven structural proteins (VP1 to VP7) and three nonstructural proteins (NS1, NS2, and NS3/NS3a) (5,6,8,9,11). Twenty-four BTV serotypes have thus far been recognized in the world, and a putative 25th serotype has recently been proposed (BTV25) (2). Seven BTV serotypes (BTV1, BTV2, BTV3, BTV4, BTV12, BTV15, and BTV16) have been isolated in China since the isolation of the first Chinese BTV strain in Yunnan Province in 1979 (3, 10, 12). Worldwide, there has been no report of the full genomic sequence of the BTV16 strain. It is necessary to acquire and analyze the sequence of the complete genome of a BTV16 strain to study the molecular features of this serotype.In the present study, we report the full genomic sequence of the serotype 16 BTV strain BN96/16, which was isolated from the blood of a sheep that showed obvious signs of bluetongue disease in Yunnan Province, China, in 1996. Viral dsRNA from BN96/16 was prepared according to the protocol described by Attoui et al. (1). Fulllength cDNA copies of each genomic segment were synthesized in a sequence-independent manner using the full-length amplification of cDNAs (FLAC) method, and subsequent rounds of PCR amplification were performed as described by Maan et al. (4). The 10 PCR products corresponding to each of the genomic segments of BN96/16 were purified and cloned into the pEASY-Blunt clone vector (Transgen Biotech, China). The cloned inserts were then sequenced by the Invitrogen Company to determine the full genomic sequence of BN96/16.The nucleotide sequences of the 10 BN96/16 segments were determined. The nucleotide lengths of BN96/16 segments 1 through 10 are as follows: segment 1, 3,944 bp; segment 2, 2,935 bp; segment 3, 2,772 bp; segment 4, 1,981 bp; segment 5, 1,763 bp; segment 6, 1,637 bp; segment 7, 1,156 bp; segment 8, 1,125 bp; segment 9, 1,052 bp; and segment 10, 822 bp. The amino acid (aa) lengths of the seven structural proteins of BN96/16 (VP1 to VP7) are as follows: VP1, 1,302 aa; VP2, 959 aa; VP3, 901 aa; VP4, 644 aa; VP5, 526 aa; VP6, 330 aa; and VP7, 349 aa. The amino acid lengths of the three nonstructural proteins of BN96/16 (NS1, NS2, and NS3/NS3a) are as follows: NS1, 552 aa; NS2, 354 aa; and NS3/NS3a, 229/216 aa.The data presented here are the first to report the complete sequence of the 10 genomic segments of a strain of BTV16. It is h...

Complete Genome Sequence of Bluetongue Virus Serotype 16 of Goat Origin from India

“…The sequences were assembled with the Newbler assembler (454 Life Sciences). The GS reference mapper was used to generate contigs (with BTV16 from China used as a reference genome (13)), and the resulting contigs were used for aligning it to the reference sequence (13), which generated a consensus sequence of sample cDNA libraries. The gaps that remained at the ends were filled by Sanger's sequencing.…”

“…The gaps that remained at the ends were filled by Sanger's sequencing. The genome annotation and comparative analysis of the genome were carried out with published genome of BTV16 from China (13).…”

“…The total size in nucleotide sequences of the 10 genome segments (seg1 to seg10) of the strain Goat/10/Ind/ABT/Hisar was determined as 19,185 bp, and the sizes of individual segments in base pairs are 3, 944, 2,929, 2,772, 1,981, 1,765, 1,637, 1,156, 1,125, 1,053, and The only previous report on the BTV16 full genome sequence is from China (13). The second report of BTV16 strain Goat/10/ Ind/ABT/Hisar, from India, would help to understand the origin and spread of the virus to various geographical regions across countries.…”

Complete Genome Sequence Analysis of a Reference Strain of Bluetongue Virus Serotype 16

“…Previously, two BTV-16 field strains from China and Australia have been fully sequenced (3,16). All genome segments of the BTV-16 reference strain (RSArrrr/16) showed Ͼ99% sequence identity to the BTV-16 vaccine strain (RSAvvvv/16) that was derived from it and to the Chinese BTV-16 (strain BN96/16) isolated from a sheep in Yunnan province during 1996 (16).…”

Complete Genomic Sequence of Bluetongue Virus Serotype 1 from China

“…Viral double-stranded RNA (dsRNA) preparation, full-length cDNA synthesis, PCR amplification, cloning, and sequencing of SZ97/1 were performed according to the protocol described by Yang et al (11).…”