SummaryCalpastatin (CAST) is a specific inhibitor of calcium-dependent neutral protease μ-calpain found in mammalian tissues. The genetic variants in the bovine CAST gene were analysed by using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The animal material of the study consisted of 132 bulls and heifers in Turkish Grey steppe cattle and Turkish Grey steppe x Brown Swiss crossbreds. C and G alleles are characterized by CAST/RsaI polymorphism were detected in the sample of animals were studied. Allele frequency C was significantly frequent in the crossbred group compared to the pure Turkish Grey steppe animals (P<0.05). In the total samples of animals, the average allele frequency C was 56.1%. Genotypic frequencies were estimated as 0.257, 0.499 and 0.243 in the purebred Turkish Grey steppe, and 0.388, 0.470 and 0.142 in the Turkish Grey steppe x Brown Swiss crossbred population for genotypes CC, CG and GG, respectively. As a result, genotypic distributions were equilibrium in both pure and crossbred examples of Turkish Grey steppe cattle.
Although most consumers are sensitive about the origin of the meat they consume, adulteration of meat products is not uncommon. For this reason, the development of reliable methods for animal species identification in meat products is an important research priority for food scientists. Species-specific protein-and DNA-identification methods generally used for this purpose. ELISA and protein electrophoresis are used for protein, PCR is used for species-specific DNA identification. Because DNA is known to be more resistant to processing than protein, PCR methods are generally considered as more sensitive for processed foods. However, processing conditions may also degrade DNA resulting in decreased DNA quality and yield. In this study, the individual and combined effects of heat treatment and low pH on the identification of animal species in meat products by PCR were evaluated. Beef sausage mixtures containing two different amounts of meat from a secondary species (either poultry, pork, or horse) were prepared and were subjected to heat treatment (65°C, 85°C, and 121°C) and pH adjustment (5.2 and 6.2). PCR screening for the four animal species was performed using DNA extracts of these meat samples. The results showed that, the combined effect of high temperature and low pH significantly affects the detection limit of the PCR method. Nevertheless, even low levels of adulteration can still be detected fallowing heat treatment. Keywords: Animal species identification, PCR, Heat processing, DNA degradation, Low pH Proses Koşullarının Et Ürünlerinde Hayvan Türünün PCR İle Teşhisine Etkisi ÖzetBirçok tüketicinin tükettiği etin orijini konusunda hassasiyetinin bulunmasına karşın, et ürünlerinde yapılan hileler oldukça sıktır. Bu sebeple, çeşitli hayvan türlerinin et ürünlerinde tespit edilmesinde kullanılacak güvenilir metotların geliştirilmesi, gıda bilimcilerinin öncelikli çalışma konularındandır. Bu amaçla, türe spesifik protein yada DNA'nın tanımlanması yöntemleri kullanılır. Protein tanımlanması için ELISA ve protein elektroforezi kullanılırken, türe spesifik DNA tanımlanmasında PCR yönteminden yararlanılır. DNA'nın proses koşullarına daha dayanıklı olduğu bilindiğinden, PCR işlem görmüş ürünlerde daha hassas olarak kabul edilmektedir. Buna karşın, proses koşulları DNA üzerine de yıkımlandırıcı etki göstererek, DNA kalite ve miktarını zayıflatabilir ve böylece PCR metotlarının işlenmiş ürünlerde kullanımını sınırlandırabilir. Bu çalışma ile ısıl işlem ve düşük pH'nın bağımsız ve kombine etkisinin et ürünlerinde hayvan türünün PCR ile teşhisine olan etkisini incelenmiştir. Bu amaçla, iki farklı seviyede ikincil bir türe ait (tavuk, domuz yada at) et katılmış olan deneysel sığır sosis karışımları yaygın olarak kullanılan proses şartlarını temsil etmek üzere ısıl işleme (65°C, 85°C, and 121°C) ve pH ayarlamasına (5.2 and 6.2) tabi tutuldu. Bu karışımların DNA ektraktlarının, PCR ile tür tayini testleri gerçekleştirildi. Elde edilen sonuçlar, düşük pH ve ısıl işlemin kombine etkisinin tespit limiti üzerine olduk...
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