PSIV-7 Targeting Variation in the Mitochondrial Genomes of Bighorn and Domestic Sheep.

ABSTRACT: Abstract The mitochondrial genome has been used in numerous species to discern evolutionary patterns, phylogeny, and speciation. Previous studies including mitochondrial genome assemblies in bighorn and domestic sheep indicate variation between these two species. This variation can be used to determine whether animal tissue originates from bighorn or domestic species. In this study, we designed and validated an assay that uses amplification of two targets in the mitochondrial genome followed by short read sequencing to identify single nucleotide polymorphisms (SNPs) in bighorn and domestic sheep. Tissue samples were acquired from two Rocky Mountain bighorn sheep and two domestic sheep (Suffolk and Icelandic breeds), and mitochondrial DNA was extracted. A target designed on the Cytochrome B (CYTB) gene (266 base pairs in length) and a target on the NADH dehydrogenase subunit 6 (ND6) gene (328 base pairs in length) were amplified using PCR and sequenced with an Illumina Mi-Seq. Sequences were aligned to Ovis aries Oar_v4.0 mitochondrial reference (NC_001941.1) and Ovis canadensis mitochondrial reference (JN181255), and variants were called using CLC Genomics Workbench v11.0 with sequence depth of over 40X. Bighorn sheep sequences aligned to the domestic mitochondrial reference displayed 17 SNPs in each target, with a total of 34 SNPs and 94% sequence identity, while domestic sequences displayed 0–2 SNPs for both targets and 99–100% sequence identity. Bighorn sheep sequences aligned to the bighorn mitochondrial reference displayed 0 variants and 100% sequence identity, while domestic sheep sequences displayed 16–17 SNPs in each target, with 33–34 total SNPs identified and 94% sequence identity. The assay designed in this study and variants identified can be used to determine whether samples are of Ovis aries or Ovis canadensis origin. Further, these data provide additional insight into mitochondrial genome variation between two Ovisspecies; one domestic and one wild.

SUBMITTER: Davenport K 

PROVIDER: S-EPMC6285092 | biostudies-literature | 2018 Dec

REPOSITORIES: biostudies-literature