ECE2022 Eposter Presentations Diabetes, Obesity, Metabolism and Nutrition (318 abstracts)
Development of THE allele-specific PCR method for studying insulin gene rs689 polymorphism
Talat Saatov 1 , Zafar Ibragimov 1 , Timur Alimov 2 , Elvira Ibragimova 1 , Tokhir Ishankhodjaev 1 , Zulaykho Shamansurova 1,3 , Kha Karimov 2 , Shakhnoza Azimova 4 , Kodirjon Boboev 2 & Anton Makhnyov 4
1Institute of Biophysics and Biochemistry under Mirzo Ulugbek National University of Uzbekistan, Tashkent, Uzbekistan; 2Specialized Scientific-Practical Medical Center of Hematology, Uzbekistan Public Healthcare Ministry, Tashkent, Uzbekistan; 3Tashkent Pediatric Medical Institute, Uzbekistan Ministry of Education, Tashkent, Uzbekistan; 4Academician S.U. Yunusov Institute of Chemistry of Plant Substances, Uzbekistan Academy of Sciences, Tashkent, Uzbekistan
Background: Insulin gene (INS) is known to be responsible for production of insulin by the pancreatic β-cells. It includes variable number of tandem repeats (VNTR) of oligonucleotide consequence (ACAGGGGT (G/C) (T/C) GGGG) in the promoter region (Bell et al, 1981,1982); being the gene involved in the susceptibility to various pathologies. The allele-specific PCR allows direct diagnosis of some genetically determined disorders is rather simple and precise. However, by today, there have been no data on the use of the allele-specific PCR for the INS gene rs689 polymorphism.
Aim: The work was initiated to develop a protocol for simple, quick and precise PCR method to determine INS gene rs689 polymorphism.
Materials and methods: To study INS gene rs689 polymorphism, we used a modified method of allele-specific PCR. Original design of primers was performed by means of bioinformation analysis of NCBL database, Genome Browser with BioEdit app. The genotyping was performed using Applied Systems-2720 programmed thermocycler.
Results: To pick up the INS gene nucleotide sequence from the NCBL database is the first step of the procedure. Oligoprimers was synthetized at the Academician S.U. Yunusov Institute of Chemistry of Plant Substances, Uzbekistan Academy of Sciences. The INS gene polymorphic region (rs689) was amplified by means of allele-specific PRC. The primers synthetized as the result of the step were the basis for development of novel test-system intended for determination of INS gene rs689 polymorphism by allele-specific PCR. The efficacy of the method was assessed in the studies in the frames of research grant for patients with diabetes mellitus.
Conclusion: The modified protocol based on the allele-specific PCR intended for study on the INS gene rs689 polymorphism was developed. The protocol can be used in study on various pathologies in diabetes mellitus, insulin resistance, polycystic ovarian syndrome, adenocarcinoma esophagogastric junction, neurodegenerative disorders, etc.
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Endocrine Abstracts
ISSN 1470-3947 (print) | ISSN 1479-6848 (online)
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