Sheep mitochondrial heteroplasmy arises from tandem motifs and unspecific PCR amplification

Abstract

The mitochondrial DNA control region (D-loop) is a widely used molecular marker in evolutionary and phylogeographic research. However, the occurrence of heteroplasmy of the D-loop region within individuals has rarely been investigated. In this study, a total of 85 Chinese sheep were used to amplify a partial D-loop region, and 15 heteroplasmic animals (17.64%) were identified. A comparative analysis of the PCR amplification and cloning of the D-loop sequences from the heteroplasmic samples revealed most of the sequencing profile from the heteroplasmic regions started at the beginning of a 75-bp random repeat motif. In addition, a total of 22 nonsyngeneic sequences with a D-loop were found in 61 of the clones obtained from the 4 random heteroplasmic and 3 homozygote animals, and their genomic locations were compared for homology. In summary, the D-Loop sequencing profiles appear to be heteroplasmic and could arise from tandem repeat motifs and unspecific replication during PCR amplification; however, they are not likely due to the presence of multiple mitochondrial genomes within an individual.

Keywords:

• Heteroplasmy
• mtDNA control region
• tandem repeats
• Chinese sheep

Disclosure statement

The authors report no conflicts of interest.

Additional information

Funding

This work was supported by the Fundamental Research Funds for the Central Universities (No. SWU114023), National Natural Science Foundation of China (No. 31601913).