Sequencing the complete mitochondrial genome of Eimeria mitis strain USDA 50 (Apicomplexa: Eimeriidae) suggests conserve
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ORIGINAL PAPER
Sequencing the complete mitochondrial genome of Eimeria mitis strain USDA 50 (Apicomplexa: Eimeriidae) suggests conserved start positions for mtCOI- and mtCOIII-coding regions M. E. Ogedengbe & M. A. Hafeez & J. R. Barta
Received: 8 August 2013 / Accepted: 25 August 2013 / Published online: 8 September 2013 # Springer-Verlag Berlin Heidelberg 2013
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 nearcomplete 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.
Introduction Coccidiosis occurs wherever chickens are reared (Dalloul and Lillehoj 2006). The disease frequently presents as simultaneous infections with multiple Eimeria species (Apicomplexa: Eimeriidae). The biology and relationships have been described M. E. Ogedengbe : M. A. Hafeez : J. R. Barta (*) Department of Pathobiology, Ontario Veterinary College, University of Guelph, 50 Stone Road East, Guelph, ON N1G 2W1, Canada e-mail: [email protected]
for the Eimeria species involved in the disease: Eimeria tenella, Eimeria necatrix, Eimeria maxima, Eimeria praecox, Eimeria mitis, Eimeria brunetti and Eimeria acervulina (Shirley et al. 1983; Reid and Long 1979; Costa et al. 2001; Barta et al 1997). Global economic impact of coccidiosis in chickens is estimated at US$2.4–3 billion per annum (McDougald and Reid 1997; Fernandez et al. 2003; Bera et al 2010). Like many members of the phylum Apicomplexa, Eimeria species possess a nuclear genome and two extra nuclear genomes: ~35-kilobase (kb) apicoplast genome (nonphotosynthetic plastid) and ~6-kb mitochondrial (mt) genome (Wilson and Williamson 1997; Feagin 1994, 2000). Although mt functionality is widely conserved, gene content, size, arrangement and gene expression vary dramatically among eukaryotes (Gray et al. 1999, 2001). Mitochondrial genome size ranges from more than 360 kb in the flowering plant Arabidopsis to only ~6 kb in Eimeria and related apicomplexan parasites (Gray et al. 1999). The structure and gene arrangement of apicomplexan mt genomes is highly variable. Apicomplexan mt genomes have been recorded as linear concatemers of a basic mt
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