Department of Pathobiology, Ontario Veterinary College, University of Guelph, 50 Stone Road East, Guelph, ON, N1G 2W1, Canada.
Parasitol Res. 2013 Dec;112(12):4129-36. doi: 10.1007/s00436-013-3604-z. Epub 2013 Sep 8.
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.
从单卵囊衍生的柔嫩艾美耳球虫 USDA50 单一系中获得了四个完整的线粒体 (mt) 序列（三个来自克隆全基因组 PCR 产物，一个来自直接测序全基因组 PCR 产物）。mt 基因组长 6408bp，包含三个基因（CytB、细胞色素 c 氧化酶亚基 I（COI）和细胞色素 c 氧化酶亚基 III（COIII））和许多 rDNA 片段（大亚基 rDNA13、小亚基 rDNA10）；组织与其他艾美耳球虫 mt 基因组相同。所有艾美耳球虫 mt 基因组的 COI 和 COIII 都建议使用保守的起始密码子位置；这些起始密码子位置存在且可能在相关的顶复门寄生虫中也保守。在接近完整的 mt 基因组的三个单独克隆 PCR 产物中，与直接测序的 mt 基因组相比，有 26 个核苷酸差异（总共）。这些变化似乎是 PCR 过程中的碱基误掺入。与克隆和随后的测序相比，直接测序长 PCR 扩增产物可能更有可能生成准确的 mt 基因组序列。
