Genome Sequences of Rotavirus A Strains Ty-1 and Ty-3, Isolated from Turkeys in Ireland in 1979

To obtain complete genome sequences of turkey rotavirus A strains Ty-1 and Ty-3, we sequenced the gene segments that had not been decoded previously. The genotype constellations of the respective strains were determined to be G17-P[38]-I4-R4-C4-M4-A16-N4-T4-E4-H4 and G7-P[35]-I4-R4-C4-M4-A16-N4-T4-E11-H14. Notably, their VP4 and NSP5 genes were classified into novel genotypes.

To understand the ecology and evolution of RVA, it is necessary to accumulate genetic information on RVA strains circulating in various host species. However, while numerous human and animal RVAs have been genetically analyzed at the whole-genome level, avian RVAs have not been extensively studied: currently, complete genome sequences of only six avian RVA strains have been determined (references 4-7 and unpublished data).
Avian RVA strains Ty-1 and Ty-3 were isolated from turkeys in Ireland in 1979 (8) and have been considered as representative avian RVAs. However, their complete genome sequences have not been determined yet: only the VP6-encoding segment of both strains and the NSP2-and VP7-encoding segments of the Ty-1 strain were previously sequenced (9)(10)(11). To obtain complete genome sequences of Ty-1 and Ty-3, we determined the gene segments that had not been decoded previously.
Viral RNA was extracted using ISOGEN-LS (Nippon gene). Libraries for next-generation sequencing (NGS) were constructed using the NEBNext Ultra RNA library prep kit for Illumina version 2.0 (New England Biolabs) according to the manufacturer's guidelines. NGS was carried out on the MiSeq bench-top sequencer (Illumina). Contigs were assembled from the obtained sequence using the de novo assembly command with default parameters in CLC Genomics Workbench 6.0 (CLC bio). The nucleotide sequences of 5= and 3= untranslated regions (UTRs) were determined by a 3= rapid amplification of the cDNA end method (3= RACE) (4). The genotypes were determined according to the guidelines of the RCWG (3) using the online genotyping tool RotaC (http://rotac.regatools.be) (12) and BLAST (http://blast .ncbi.nlm.nih.gov/Blast.cgi).
Nucleotide sequence accession numbers. The nucleotide sequences determined in this study have been deposited in GenBank under the accession numbers LC088107 to LC088124.

ACKNOWLEDGMENT
This work was supported by JSPS KAKENHI grant 26292148.

FUNDING INFORMATION
The Japan Society for the Promotion of Science (JSPS) provided funding to Makoto Sugiyama under grant number 26292148.