Development of a method for the identification of rs6265 polymorphism in the human brain neurotrophic factor gene

Abstract

The analysis and generalization of data from the literature sources characterizing the structural organization, regulation of expression, and functional activity of the neurotrophic factor of the human brain, which is one of the most common regulators of biological processes in the nervous system, is carried out. The results of numerous studies demonstrating the association of the brain-derived neurotrophic factor (BDNF) gene with the pathophysiology of the affective disorders are noted, and its contribution to the development of neuroplasticity is confirmed. The results of the design of a pair of primers and adaptation of the amplification reaction of the BDNF region with a length of 433 nucleotide pairs containing the polymorphic locus RS6265 are presented. Restriction endonuclease was selected. The sequence of primers, their localization, and correlation with the restriction site provided the separation of alternative alleles necessary for the successful identification of this marker. The use of the proposed technique made it possible to uniquely identify the genotype in 38 examined whole blood samples and identify the rare allelic variants. Also, the frequency of polymorphic variants of RS6265 of the BDNF gene was established in all the samples. There was an increase in the proportion of genotypes G/A and A/A of the RS6265 polymorphism of the BDNF gene in the group of examined systematically exposed to extreme factors. Identification of people with a rare A/A genotype of the RS6265 polymorphic locus of the BDNF gene is of a great importance for the monitoring system of long-term potentiation processes leading to the development of neuropsychic pathology. The possibility of implementing this method of genotyping in a typical laboratory using a polymerase chain reaction is proved. The proposed version of the polymerase chain reaction with the subsequent analysis of the polymorphism of the lengths of restriction fragments could be used as a fast, inexpensive, and reliable system for identifying single-nucleotide genetic polymorphisms.

Keywords

molecular genetic markers / polymerase chain reaction / brain neurotrophic factor / point genotyping / single nucleotide polymorphism / neurogenesis / stress / neuroplasticity / RS6265 polymorphism identification technique

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RIGHTS & PERMISSIONS

Kutelev G.G., Krivoruchko A.B., Trandina A.E., Morozova N.E., Cherkashin D.V., Ivanov A.M., Ovchinnikov D.V., Glushakov R.I.