Українська версія | English version
THE INTERDEPARTMENTAL SUBJECT SCIENTIFIC DIGEST PIG BREEDING

and agroindustrial production

TECHNIQUE OF PCR ANALYSIS OF MT2A GENES IN DIFFERENT ANIMAL SPECIES


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UDC: [575.113+575.22]:[636.4+636.028]:577.213.3

DOI: 10.37143/2786-7730-2024-3(81)2

REFERENCIS АРА style:Saienko, A. M., Peka, M. Y., Bolotova, Yu. S., Lobchenko, O. V., Korinnyi, S. M., & Balatsky, V. M. (2024). Tekhnika PLR-analizu heniv MT2A u riznykh vydiv tvaryn [Technique of PCR analysis of MT2A genes in different animal species]. Svynarstvo i ahropromyslove vyrobnytstvo [Pig Breeding and Agroindustrial Production]. Poltava, 3(81), 22-28 [in Ukrainian]. doi: 10.37143/2786-7730-2024-3(81)2

A. M. Saienko candidate of agricultural sciences, senior researcher, performing the duties of the head lab genetics
ORCID:https://orcid.org/0000-0002-0527-5367
E-mail:saenko_artem@meta.ua
Institute of Pig Breeding and agroindustrial production NAAS, Shvedska Mohyla Str., 1, Poltava, Ukraine, 36013
M. Y. Peka researcher lab genetics
ORCID:https://orcid.org/0000-0003-0612-1164
E-mail:pekapoltava@gmail.com
Institute of Pig Breeding and agroindustrial production NAAS, Shvedska Mohyla Str., 1, Poltava, Ukraine, 36013
Yu. S. Bolotova junior researcher of the lab. genetics
ORCID:https://orcid.org/0009-0003-2526-1776
E-mail:bolotovakharkov@gmail.com
Institute of Pig Breeding and agroindustrial production NAAS, Shvedska Mohyla Str., 1, Poltava, Ukraine, 36013
O. V. Lobchenko laboratory assistant
ORCID:https://orcid.org/0009-0006-8662-2012
E-mail:oleksandra.lobchenko@gmail.com
Institute of Pig Breeding and agroindustrial production NAAS, Shvedska Mohyla Str., 1, Poltava, Ukraine, 36013
S.M Korinnyi candidate of agricultural sciences senior researcher, senior researcher,lab genetics
ORCID:https://orcid.org/0000-0002-1649-3079
E-mail:korinny_sergey@ukr.net
Institute of Pig Breeding and agroindustrial production NAAS, Shvedska Mohyla Str., 1, Poltava, Ukraine, 36013
V. M. Balatsky doctor of agricultural sciences, senior researcher, professor, chief researcher of the lab. genetics
ORCID:https://orcid.org/0000-0002-6034-3852
E-mail:vnbalatsky@gmail.com
Institute of Pig Breeding and agroindustrial production NAAS, Shvedska Mohyla Str., 1, Poltava, Ukraine, 36013

Abstract

Metallothioneins are a fam ily o f low-molecular-weight cysteine-rich proteins that participate in the metabolism o f essential metals and protect against heavy metal toxicity. Metallothioneins are represented by different isoforms in many eukaryotic and prokaryotic species. The promoter regions o f metallothionein genes contain sequences o f elements that regulate expression in response to metals (MRE), which makes it an urgent task to investigate polymorphisms in the promoter regions o f metallothionein genes. Objective. To develop and optimize the technique o f PCR analysis o f orthologous metallothionein genes in various biological species using the example o f the MT2A isoform o f rats and pigs.Methods. Analysis o f the primary structure o f MT2A isoform genes was performed using the NCBI and Ensembl databases. The design ofprimers fo r PCR analysis was carried out using the Primer3 program. Biomaterial samples were used for the study: livers o f Wistar rats and blood o f large white pigs o f intrabreed type ULW-1. DNA was isolatedfrom rat liver using the NeoPrep DNA Magnet plant DNA kit, and from pig blood using Chelex 100 reagent. Amplification o f DNA fragments was carried out using PCR followed by electrophoretic separation o f the amplicons in a 2 % agarose gel. Results. In the course o f the study, the primary structure o f the MT2A genes o f rats and pigs was analyzed. Primers were designed to cover the promoter regions o f the MT2A genes o f these two animal species. The expected sizes o f the amplicons are 583 bp and 512 bp fo r rats andpigs, respectively. The conditions fo r the synthesis o f PCR amplicons were selected, the optimality o f which is confirmed by the presence o f the corresponding PCR amplicons on the electrophoregram. Conclusions. The developed PCR analysis system fo r the MT2A genes o f rats and pigs allows to study polymorphisms in the promoter regions that contain M RE sites. Considering the importance o f MRE sequences fo r the regulation o f metallothioneins’ expression under the influence o f metals, as well as the participation o f metallothioneins in various physiological and pathological processes, it can be expected that polymorphisms in the studied areas may have associations with adaptation potential, resistance to heavy metals, economic qualities o f organisms, etc. In the future, research involving the MT2A gene as a candidate fo r the development o f molecular genetic markers and their use in the practice o f marker-associated selection is promising

Key words: metallothioneins, polymorphism, Rattus norvegicus, Sus scrofa domesticus, polymerase chain reaction, promoter.

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