Genotypic structure of micropopulations of domestic pigs at the locus of the ryanodine RYR1 receptor

T. M. Ryk
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Abstract

An important problem in transplant medicine is the lack of organs and tissues for human transplantation. One of the promising areas of its solution today is the use of organs and tissues of animals — xenotransplantation. Pigs for medical and biological needs must be stress-resistant and adapted to the conditions of keeping and feeding within the vivarium. Stress syndrome in pigs is caused by a point mutation that replaces 1843CT in exon 17 of the ryanodine RYR1 receptor gene in a homozygous state. The aim of the study was to examine the genetic structure of individual pig populations at the locus of the RYR1 gene responsible for hypersensitivity. DNA testing of 102 pigs of Poltava meat, Myrhorod, Great White, Landrace and Pietren breeds was performed. The studies were performed on DNA samples obtained from pig blood. DNA was isolated using Chelex 100 reagent. Studies of the RYR1 gene fragment were performed by restriction fragment length polymorphism (RFPR) by polymerase chain reaction (PCR). Two alleles of the ryanodine RYR1 receptor gene (n and N) and three genotypes (NN, Nn, nn) were identified in the studied animals. A molecular study of pigs of different breeds revealed a polymorphism of the RYR1 gene. The number of animals carrying the recessive allele n of the ryanodine receptor gene RYR1, which is responsible for the sensitivity of pigs to stress factors, varied considerably: from its complete absence in the Vietnamese Pot-bellied, Ukrainian meat, Large White and Ukrainian Spotted Steppe breeds up to 100% of animals of homozygous RYR1nn genotype in the Piétrain breed and 50% of Landrace individuals with the heterozygous RYR1Nn genotype The mutant RYR1n allele was found in animals of the Piétrain breed with a 100% frequency. Heterozygous genotype RYR1Nn was found in pigs of Poltava Meat (10%), Landrace (50%), Myrhorod (15%) Ukrainian Spotted Steppe (50%). Homozygous RYR1NN genotype was detected in all Large White, Vietnamese Pot-bellied, and Ukrainian Meat pigs studied, indicating the absence of stress syndrome. It has been established that pigs of the Great White, Vietnamese Pot-bellied, and Ukrainian Meat breeds are preferred for breeding for biomedical purposes. Breeding of Piétrain pigs and any combinations with this breed makes it impossible to use such animals for biomedical experimental work.
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家猪ryanodine RYR1受体位点微群的基因型结构
移植医学的一个重要问题是人体移植器官和组织的缺乏。其解决方案的一个有前途的领域是利用动物的器官和组织——异种移植。用于医疗和生物需求的猪必须具有抗应激能力,并适应在舍内饲养和喂养的条件。猪的应激综合征是由一个点突变引起的,该突变取代了ryanodine RYR1受体基因17外显子的1843CT,处于纯合状态。该研究的目的是检查个体猪群中导致过敏的RYR1基因位点的遗传结构。对102头波尔塔瓦肉猪、默荷罗德猪、大白猪、长白猪和彼得伦猪进行了DNA检测。这些研究是在从猪血液中获得的DNA样本上进行的。采用Chelex 100试剂分离DNA。采用聚合酶链反应(PCR)对RYR1基因片段进行限制性片段长度多态性(RFPR)研究。在研究动物中鉴定出2个ryanodine RYR1受体基因等位基因(n和n)和3个基因型(NN、NN、NN)。对不同品种猪的分子研究揭示了RYR1基因的多态性。携带ryanodine受体基因RYR1隐性等位基因n的动物数量差异很大,该基因负责猪对应激因素的敏感性。从越南大肚、乌克兰肉牛、大白和乌克兰斑点草原品种中完全没有RYR1n等位基因,到100%的纯合RYR1nn基因型动物和50%的长白猪个体具有杂合RYR1nn基因型,在pi15.3品种的动物中发现突变RYR1n等位基因,频率为100%。在波尔塔瓦肉猪(10%)、长白猪(50%)、Myrhorod猪(15%)和乌克兰斑点草原猪(50%)中发现杂合基因型RYR1Nn。在研究的所有大白猪、越南大肚猪和乌克兰肉猪中检测到纯合子RYR1NN基因型,表明没有应激综合征。已经确定,大白猪、越南大肚猪和乌克兰肉猪品种更适合用于生物医学目的的育种。pisamettrain猪的繁殖以及与该品种的任何组合都不可能将此类动物用于生物医学实验工作。
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