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Analysis of CHD Gene Polymorphism as a Model Object for Molecular Sexing of Eurasian Eagle-Owl (Bubo bubo)

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Abstract

CHD gene polymorphism as a model object for molecular sex determination in Bubo bubo (Eurasian eagle-owl) was studied. Classical PCR methods for amplification of target regions of the CHD gene using P2/P8, 2550F/2718R, and 123L/1272H marker systems and restriction (PCR-RFLP) and heteroduplex methodological approaches were analyzed. It was shown that the use of the P2/P8 marker system within classical PCR is efficient for avian sexing only under electrophoretic separation of the fragments in polyacrylamide gel. The application of 2550F/2718R and 123L/127H markers demonstrated their lack of efficiency both in agarose and polyacrylamide gels because it was not possible to differentiate between genotypes CHDZZ and CHDZW. The presence of a polymorphic restriction site for HaeIII in CHDZ and CHDW fragments amplified with primers P2/P8 was proven. No polymorphic site was detected for DdeI restriction endonuclease. High efficiency of the heteroduplex analysis based on the P2/P8 markers for sexing individuals under electrophoretic separation of the amplified fragments both in agarose and polyacrylamide gels was proven. In case of experimental samples containing CHDZ and CHDW amplicons, additional fragments of heteroduplex DNA were found, the presence of which makes it possible to accurately determine the genotype of an avian.

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The research was not funded by third-party commercial organizations.

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Authors and Affiliations

  1. National University of Life and Environmental Sciences of Ukraine, 03041, Kyiv, Ukraine

    R. O. Kulibaba

  2. Institute of Animal Science, National Academy of Agrarian Sciences of Ukraine, 61026, Kharkiv, Ukraine

    Yu. V. Liashenko

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  1. R. O. Kulibaba
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Correspondence to R. O. Kulibaba.

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Conflict of interests. The authors declare that they have no conflicts of interest.

Statement on the welfare of animals. Studies were carried out using Bubo bubo feathers as a source of biological material without direct manipulations concerning birds.

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Translated by K. Lazarev

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Kulibaba, R.O., Liashenko, Y.V. Analysis of CHD Gene Polymorphism as a Model Object for Molecular Sexing of Eurasian Eagle-Owl (Bubo bubo). Cytol. Genet. 55, 324–330 (2021). https://doi.org/10.3103/S0095452721040071

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