DNA MARKERS BASED ON SINGLE NUCLEOTIDE POLYMORPHISMS IN THE LEPTIN GENE

Abstract

Single-nucleotide polymorphisms within the leptin hormone gene (LEP) could potentially serve as DNA markers of productive traits in pigs. Thus, the search and development of promising LEP gene polymorphisms is one of the urgent tasks. Leptin is a protein of 167 amino acid residues and a molecular weight of 16 kDa, and the corresponding LEP gene is located on chromosome 18 in the pig genome. Leptin-mediated signaling plays a fundamental role in animal food intake and energy expenditure, thereby influencing body weight. Goal. To analyze the primary structure of the pig LEP gene, to determine single nucleotide polymorphisms, to develop a DNA typing system for the LEP gene. Methods. Blood samples from 10 Large White pigs of the intrabreed type ULW-1 sourced from the Stepne State Enterprise in Stepne village, Poltava district, Poltava region, were used for DNA typing of the LEP gene. DNA isolation from biomaterial was carried out using the NeoPrep DNA Magnet plant DNA sample preparation kit. Genotyping was planned using the PCR-RFLP technique. Results. An analysis of the primary structure of the LEP gene from the Ensembl database (ID: ENSSSCG00000040464) was conducted. Primers were designed and PCR conditions were chosen for synthesizing the LEP gene amplifier. The selected amplification conditions allow the synthesis of a 310-bp fragment. The presence of the appropriate PCR amplifier on the electrophorogram confirms the correct choice of all parameters. Separation of allelic variants of the rs1110706811 polymorphism by the PCR-RFLP method is possible using the TaqI endonuclease. Conclusions. It is expected that the developed DNA-typing technique for the LEP gene will be used to analyze its polymorphism in pig gropus for which parameters of productive qualities have been determined. Considering that different SNPs of the LEP gene associate with productive qualities of pigs and physicochemical parameters of pork meat, it can be expected that the studied polymorphism rs1110706811 of the LEP gene could potentially have a relationship with the these parameters.