Sequencing the complete mitochondrial genome of Eimeria mitis strain USDA 50 (Apicomplexa: Eimeriidae) suggests conserved start positions for mtCOI- and mtCOIII-coding regions

Abstract

Four complete mitochondrial (mt) sequences from a single-oocyst-derived line of Eimeria mitis USDA 50 were obtained (three from cloned whole-genome PCR products, one from directly sequenced whole-genome PCR product). The mt genome is 6,408 bp long with three genes (CytB, cytochrome c oxidase subunit I (COI) and cytochrome c oxidase subunit III (COIII)) and many rDNA fragments (large subunit rDNA 13, small subunit rDNA 10); organisation was identical to other Eimeria sp. mt genomes. Conserved start codon positions for both COI and COIII are suggested for all Eimeria mt genomes; these start codon positions exist and may also be conserved, in related apicomplexan parasites. Within the three separate cloned PCR products of near-complete mt genomes, there were 26 nucleotide differences (collectively) compared to the directly sequenced mt genome. These changes appear to be base misincorporations during PCR. Direct sequencing of long PCR amplification products may be more likely to generate accurate mt genomic sequences than cloning and subsequent sequencing.

Complete mitochondrial genome sequences from five Eimeria species (Apicomplexa; Coccidia; Eimeriidae) infecting domestic turkeys

“…Purification of oocysts and genomic DNA extraction was carried out as previously described by Ogedengbe et al [9,24]. Mitochondrial whole genome amplification for all five Eimeria species was initiated using two sets of specific primers that generated overlapping PCR fragments: 1) Cocci_MT-WG-F (5′-TACACCTAGCCAACACGAT-3′) and Cocci_MT-WG-R (5′-GCAGCTGTAGATGGATGCTT-3′); and, 2) Inv_COI_262R (5′-AAWGCGGCATCRTAGAATTG-3′) and Inv_COI_461F (5′-CTAGCYATGGGATGTATTACTG-3′).…”

“…The specific LSU and SSU rDNA fragments found in the mt genome of apicomplexan parasites differ among distantly related parasites. Unlike many eukaryotic mt genomes, apicomplexan mt genomes do not encode 5S rRNA or tRNAs [20-24]. …”

The Complete Mitochondrial Genome Sequence of Hepatozoon catesbianae (Apicomplexa: Coccidia: Adeleorina), a Blood Parasite of the Green Frog, Lithobates (Formerly Rana) clamitans

“…by Lin et al, 2011;Ogedengbe et al, 2013). Generating high-quality sequence required the amplification of 3 smaller, overlapping fragments of the mitrochondrial genome of H. catesbianae.…”

“…The physical structure of the mitochondrial genome copies of H. catesbianae remains unknown, and examining related apicomplexan parasites provides little guidance. Among apicomplexan protists, mitochondrial genome structures are highly diverse, with several structural forms (circular, linear, or linear-concatenated) being reported (Gray et al, 2004;Hikosaka et al, 2010;Hikosaka, Nakai et al, 2011;Hikosaka, Watanabe et al, 2011;Lin et al, 2011;Feagin et al, 2012;Liu et al, 2012;Ogedengbe et al, 2013).…”

“…However, the order and orientation of the 3 protein-coding regions varies considerably among different apicomplexan groups. The genome structure is conserved among closely related parasites such as among piroplasms (Hikosaka et al, 2010), among hemosporinids (Hikosaka, Watanabe et al, 2011), and among eimeriid coccidia (Hikosaka et al, 2010;Lin et al, 2011;Ogedengbe et al, 2013). The mitochondrial genome organization of H. catesbianae (Fig.…”