English: Quantification of Apocryptovirus odysseus (Ao) in Toxoplasma gondii strains COUGAR (aka COUG), RUB and others
(A) Quantification of Ao-aligning reads amongst thirty strains of T. gondii strains and mock control, error bars indicate 2 SD.
(B) Unrooted T. gondii strain phylogeny (IQ-TREE) from transcriptome, and strain cross-contamination ruled out by transcriptome variant analysis. The original geographic isolation of strains is highlighted as North American, European, or South American, and lines are colored by the genomic T. gondii clades. A. odysseus (Ao, red arrow) was detected at high levels in two geographically and phylogenetically unrelated strains of T. gondii; RUB isolated from a symptomatic soldier in French Guiana in 1991 and COUGAR isolated from a cougar in British Columbia in 1996.
(C) RNA sequencing expression data showing normalized expression levels of Ifnb1 in murine macrophages infected with strains of T. gondii.
(D) Pairwise nucleotide sequence identity of complete coding sequences from contig 1 and 2
(E) Reverse Transcriptase (RT) PCR for Ao or GRA1, a highly expressed Toxoplasma gene used as an RNA quality control, from Ao+ (RUB and COUGAR) and Ao—control RH[delta hpt] strains of Toxoplasma grown on HFFs. Asterisks denote non-specific bands. Sanger sequencing of amplicons showed 100 % nucleotide identity to the assembled Ao RUB and Ao COUGAR, respectively.