A development of a PCR-RFLP test system for the identification of mitochondrial lines of the Pelophylax ridibundus lake frog in Kazakhstan 

Daniya A. Ualiyeva, Junior researcher of the laboratory of ornithology and herpertology, Institute of Zoology (93 al-Farabi avenue, Almaty, Kazakhstan), E-mail: daniya.2010@mail.ru
Alexander Yu. Ivanov, Candidate of biological sciences, lecturer of the sub-department of zoology and ecology, Penza State University (40 Krasnaya street, Penza, Russia), E-mail: akella58@mail.ru
Oleg A. Ermakov, Candidate of biological sciences, associate professor, associate professor of the sub-department of zoology and ecology, Penza State University (40 Krasnaya street, Penza, Russia), E-mail: oaermakov@list.ru 

Background. Molecular typing by PCR-RFLP method allows to identify the specific attribution of an organism that has a weak phenotypic difference. The main advantage of this research method is the capacity to analyze a large number of samples without applying the sequencing method. The purpose of the work is to develop a test-system for the identification the matrilines of marsh frogs of the Pelophylax ridibundus complex from Kazakhstan. Materials and methods. The analysis was based on the variability of the prima-ry structure of the mitochondrial gene the second subunit of dehydrogenase (ND2) which is a species-specific marker. Subsequently a search for marker nucleotide substitutions specific for each lineage and recognition sites for the HaeIII and TasI restriction endonucleases was conducted. Results. As a result of the research, it was confirmed that on the territory of Kazakhstan inhabiting three main forms of lake frogs where two of them are native (Balkhash, Syrdarya), and invasive Anatolian form – P. cf. bedriagae. Conclusions. The authorsʼ PCR-RLFP test technique is a straightforward and reliable tool for detecting mitochondrial lineages in the Pelophylax ridibundus complex marsh frogs and may be used successfully in mass screening investigations. 

restriction analysis, NADH dehydrogenase (ND2), mitochondrial DNA, Pelophylax ridibundus complex 

Ualiyeva D.А., Ivanov A.Yu., Ermakov О.А. A development of a PCR-RFLP test system for the identification of mitochondrial lines of the Pelophylax ridibundus lake frog in Kazakhstan. Izvestiya vysshikh uchebnykh zavedeniy. Povolzhskiy region. Estestvennye nauki = University proceedings. Volga region. Natural sciences. 2022;(1):76–84. (In Russ.). doi:10.21685/2307-9150-2022-1-7