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Length heteroplasmy of the polyC-polyT-polyC stretch in the dog mtDNA control region

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Abstract

Previously, the mitochondrial control region of 214 Belgian dogs was sequenced. Analysis of this data indicated length heteroplasmy of the polyT stretch in the polyC-polyT-polyC stretch from positions 16661 to 16674. Nine polyC-polyT-polyC haplotype combinations were observed, consisting of seven major haplotypes (highest signal intensity) combined with minor haplotypes (lower signal intensity) one T shorter than the major haplotype in all but three dogs. The longer the polyT stretch, the smaller was the difference in signal intensity between the major and minor haplotype peaks. Additional sequencing, cloning, and PCR trap experiments were performed to further study the intra-individual variation of this mitochondrial DNA (mtDNA) region. Cloning experiments demonstrated that the proportion of clones displaying the minor haplotypes also increased with the length of the polyT stretch. Clone amplification showed that in vitro polymerase errors might contribute to the length heteroplasmy of polyT stretches with at least 10 Ts. Although major and minor polyC-polyT-polyC haplotypes did not differ intra-individually within and between tissues in this study, interpretation of polyT stretch variation should be handled with care in forensic casework.

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Acknowledgments

We thank L. Cluyts for her technical assistance and NICC staff members, dog training school Koninklijke Sint-Niklase Hondenafrichtingsclub Belsele, veterinarian M. Rober, and animal shelters Dierenasiel Mechelen, De Grensstreek Rekkem, and Leiestreek Zwevegem for dog sampling. S. Verscheure is a PhD student at the University of Antwerp supported by a grant from the Belgian Science Policy Office. This work was partially conducted within the framework of the FWO Research Community W0.009.11N “Belgian Network for DNA Barcoding.”

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. National Institute of Criminalistics and Criminology, Vilvoordsesteenweg 100, 1120, Brussels, Belgium

    Sophie Verscheure & Stijn Desmyter

  2. University of Antwerp (Evolutionary Ecology Group), Groenenborgerlaan 171, 2020, Antwerp, Belgium

    Sophie Verscheure & Thierry Backeljau

  3. Royal Belgian Institute of Natural Sciences (OD “Taxonomy and Phylogeny” and JEMU), Vautierstraat 29, 1000, Brussels, Belgium

    Thierry Backeljau

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  1. Sophie Verscheure
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Correspondence to Sophie Verscheure.

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Verscheure, S., Backeljau, T. & Desmyter, S. Length heteroplasmy of the polyC-polyT-polyC stretch in the dog mtDNA control region. Int J Legal Med 129, 927–935 (2015). https://doi.org/10.1007/s00414-014-1106-x

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  • DOI: https://doi.org/10.1007/s00414-014-1106-x

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