Genetic variation of CAST gene in Local Karnobat and Karnobat merino sheep breeds

Karnobat sheep plays an important role in the development of sheep breeding in Southeastern region of Bulgaria. They are valuable source of genetic material. The aim of present experiment was to determine the allele variation of CAST gene in Local Karnobat and Karnobat Merino sheep breeds. A total of 60 blood samples were collected – 30 per breed. DNA was extracted and genotypes of all animals were identified by means of PCR-RFLP technique. The restriction reactions were accomplished by specific enzyme MspI. As expected both breeds were characterized with low level of genetic diversity due to the fact that mostly maintaining selection has been implemented. In Local Karnobat sheep breed was identified only one heterozygous individual from all 30. In Karnobat merino were identified allele M with frequency 0,97 and allele N with frequency 0,03. Genotypes MM and MN were revealed with frequencies 0,93 and 0,07, respectively. According to the statistical analysis both breeds were in HWE equilibrium.

INFLUENCE OF POLYMORPHIC GENES ON QUALITATIVE INDICATORS OF MEAT PRODUCTS

Проблема и цель. Целью данной работы являлось генотипирование животных по генам CAST и CAPN1 c выявлением частот генотипов и аллелей, а также тестирование продукции этих животных, выраженное в оценке выдержанного в течение 8 суток мяса, на показатели, составляющие нежность, сочность, органолептическую оценку вкуса. Методология. Объектом исследований являлись бычки кроссов брединского мясного типа симменталов, полученные от использования выдающихся быков-производителей канадских мясных симменталов на отечественных матках различных линий. Для осуществления полимеразной цепной реакции (ПЦР) по тиражированию участка генома, составляющего ген CAPN1316 и ген CAST2857, использовались праймеры, найденные в открытой печати и синтезированные в фирме «Синтол». Проводилась оценка мяса на нежность (сопротивление усилию на разрез прибором Уорнера-Братцлера + органолептическая оценка варёного и жареного мяса), сочность (p/h, с использованием прокалывающего электрода LoT406-M6-DXK-S7/25 + влагоудерживающая способность). Результаты. В результате показатели мяса бычков, имеющих сочетание генотипов TT* по CAST и СС* по CAPN1, имели превосходство над средними показателями общей выборки (P< 0,001); а также над имеющей наибольшее распространение в выборке группой животных с желательным генотипом СС по гену CAPN1 на 1,22 балла или 15,4 % (P< 0,01) по нежности; на 1,44 балла или 18,2 % (P< 0,05) по сочности; на 1,56 балла или 19,2 % (P< 0,001) по вкусу. Заключение. Таким образом, исследования подтвердили влияние полиморфного состояния гомозиготы СС гена CAPN1 само по себе, а также генов CAST и CAPN1 на нежность говядины в той её части, когда сочетание генотипов ТТ в гене CAST и СС в гене CAPN1 даёт положительный эффект по нежности говядины, вероятно связанный с активизацией деятельности μ-кальпаина в связи с ослаблением влияния кальпастатина как ингибитора на μ-кальпаин. Problem and purpose.The aim of this work was to genotype animals for the CAST and CAPN1 genes with identifying the frequencies of genotypes and alleles, as well as testing the products of these animals, expressed in assessing meat aged for 8 days, for indicators that make up tenderness, juiciness, and organoleptic taste assessment.Methodology. The object of research was the bulls of the breeds of the Bredy meat type of simmentals obtained from the use of outstanding bulls-producers of Canadian meat simmentals on domestic females of various lines. To carry out the polymerase chain reaction (PCR) for replicating a region of the genome constituting the CAPN1316 gene and the CAST2857 gene, primers found in the open press and synthesized by Syntol were used. The meat was evaluated for tenderness (resistance to cutting force with a Warner-Bratzler device + sensory evaluation of cooked and fried meat), juiciness (p/h, using a piercing electrode LoT406-M6-DXK-S7/25 + water-holding capacity). Results. As a result, the meat indicators of bulls with a combination of TT * genotypes for CAST and CC * for CAPN1 were superior over the average indicators of the total sample (P <0.001), as well as over the most common group of animals in the sample with the desired CC genotype for the CAPN1 gene by 1.22 points or 15.4% (P <0.01) for tenderness, 1.44 points or 18.2% (P <0.05) for juiciness, 1.56 points or 19.2% (P <0.001) for taste. Conclusion. Thus, the studies have confirmed the influence of the polymorphic state of the CC homozygote of the CAPN1 gene itself, as well as of the CAST and CAPN1 genes, on the tenderness of beef in that part of it when the combination of TT genotypes in the CAST gene and CC in the CAPN1 gene has a positive effect on the tenderness of beef, probably associated with the activation of the activity of μ-calpain due to the weakening of the effect of calpastatin as an inhibitor on μ-calpain.

Polymorphism of CAST gene in five sheep breeds in Bulgaria

CAST gene is a candidate marker that influences the intensity of growth and meat quality. The aim of this study was to investigate the genetic variability of CAST gene in five Bulgarian sheep breeds – two merino, two local and one for milk. A total of 150 ewes, belonging to these breeds were investigated for polymorphisms of CAST gene by using PCR-RFLP method. A 622 bp fragment of Ovis aries CAST gene was amplified using PCR. After restriction with endonuclease MspI two alleles were observed in Ascanian, Caucasian, Breznik and Pleven Blackhead breeds. One allele (M) and one genotype (MM) were detected in Cooper-Red Shumen breed. The highest frequency of the allele N was established in the Ascanian merino breed (0.27) followed by Caucasian merino (0.13), Breznik (0.12) and Pleven Blackhead (0.07). Two genotypes - MM and MN, were observed in Caucasian, Breznik and Pleven Blackhead breeds. The three possible genotypes were found only in Ascanian merino ewes - MM, MN and NN. The lowest frequency of the homozygous genotype MM (0.50) and the highest frequency of the heterozygous genotype MN was established in the Ascanian merino breed (0.47). The obtained results confirm that the PCR-RFLP method can be used to identify different genotypic variation of CAST gene in Bulgarian sheep breeds. The established genetic diversity in the calpastatin gene indicates that, after further associative studies, this gene may be included in the breeding programs of certain sheep breeds.

FATTY ACID COMPOSITION OF MUSCLE TISSUE LIPIDS IN YOUNG SHEEP OF DIFFERENT ALLELIC VARIANTS OF THE CAST GENE

The article presents the results of the analysis of the fatty acid composition of the lipids of the muscle tissue of sheep of diff erent genotypes of the Stavropol breed at the age of 8 months. The polymorphism of the calpastatin CAST gene was determined using PCR-PDRF.

Calpastatin (CAST) Gene Polymorphism in Tsigai and Merinoland Sheep Breeds Under Conditions of the Republic of Crimea

The article presents the results of studying the Calpastatin (CAST) gene polymorphism in Tsigai (n = 23) and Merinoland sheep breed (n = 13). The purpose of the research. The possibility of using PCR-RFLP analysis to detect the Calpastatin (CAST) gene polymorphism in Tsigai and Merinoland sheep was the primary objective of this ongoing study. Research methods. Calpastatin (CAST) gene polymorphism was analyzed by PCR-RFLP analysis using the MspI endonuclease restriction. Results. We revealed the diversity of genotypes and allelic variants of the Calpastatin (CAST) gene in Tsigai and Merinoland Breeds. It was found that M allele of the Calpastatin locus is the most common. Frequencies of MM, MN and NN genotypes were found to be 74, 4 and 22 % in Tsigai breed. Among the representatives of Merinoland sheep, the frequencies of MM and MN genotypes were 92 % and 8%, respectively. No animals with NN genotypes were found.

Polymorphism of genes of Growth Hormone (GH) and Calpastatin (CAST) in wool-and-meat producing sheep

At present, modern technologies involving the use of DNA markers, which are widely used in national breeding programs in a number of countries with developed animal husbandry are of considerable interest and have a signifi cant infl uence on improving productivity traits. A signifi cant number of genes associated with meat productivity and meat quality have been identifi ed. In sheep breeding such research has been developed only recently. Due to the growing interest in the production of young mutton and lamb, priority is given to the study of genes that control meat productivity. However, the most popular are genetic markers that are interconnected with candidate genes, whose protein products play a signifi cant role in the formation or regulation of physiological and biochemical processes. Alleles of the Growth Hormone (GH) gene and Calpastatin (CAST) gene are considered as potential markers of sheep meat productivity. In this regard, the purpose of this work was to study the polymorphism of GH and CAST genes in crossbred wool-and-meat producing sheep (1/2 Poll Dorset × 1/2 North Caucasian wool-and-meat producing breed) bred in the Stavropol Territory. The analysis of Somatotropin and Calpastatin gene polymorphism was performed by PCR-PDRF. When studying GH gene polymorphism, the genotypes AA, AB and BB have been found, the frequency of occurrence of which was 29,7; 42,8 and 27,5 %, respectively. Allele A prevailed in the studied animals – 0,786, allele B was – 0,214. The CAST gene was represented by two genotypes MM and MN (the NN genotype was absent in the animals of this group) with a frequency of 87,9 and 12,1 %, respectively. The frequency of the M allele was high at 0,94, while the n allele was low at 0,06. The results obtained during the researches indicate the establishment of a variety of allelic variants of Somatotropin and Calpastatin genes in wool-and-meat producing sheep. Further research will be aimed at studying the association of identifi ed polymorphisms with economically useful traits.