SUMMARY. The studies of CHD gene polymorphism as a model object for molecular sex determination of Bubo bubo (Eurasian eagle-owl) were carried out. The methods of PCR for amplification of the target regions of CHD gene, using P2/P8, 2550F/2718R, and 123L/1272H markers systems, restriction (PCR-RFLP) and heteroduplex methodological approaches were analyzed. It was shown that the use of P2/P8 markers system within 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 insufficient efficiency both in agarose and polyacrylamide gels, because of the impossibility 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, has been proven. No polymorphic site has been detected for DdeI restriction endonuclease. High efficiency of heteroduplex analysis, based on P2/P8 markers system, for sexing individuals under electrophoretic distribution of the amplified fragments both in agarose and polyacrylamide gels was proven. In case of experimental samples, containing CHDZ and CHDW amplicons, the additional fragments of heteroduplex DNA were formed. The presence of the latter allowed determining the genotype of an individual accurately.
Keywords: Strigidae, Bubo bubo, polymorphism, polymerase chain reaction, restriction, heteroduplexes
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