Всероссийский научно-исследовательский институт физиологии, биохимии и питания животных – филиал Федерального государственного бюджетного научного учреждения «Федеральный научный центр животноводства – ВИЖ имени академика Л.К. Эрнста»
ABSTRACT. Acyl-CoA:diacylglycerol acyltransferase 1(DGAT1) is a key enzyme in triacylglycerol synthesis. Among known allelic variants of bovine dgat1 gene, the non-synonymous mutation GC → AA at positions 10433/10434 (according to GenBank no. AJ318490 sequence) results in the nonhomologous substitution of amino acid residue 232 (A → K). Numerous studies showed a strong association of the DGAT1 K232 allele with increasing saturated fat content in milk and beef. Therefore, dgat1 was considered as a candidate gene for control of milk and beef fat contents and composition. The aim of this work was to improve the method for identification of K and A allelic variants of dgat1 gene by shortening the duration of the procedure for genotyping cattle. In the developed method, two primers flanking 279-bp fragment of dgat1 gene and two allele-specific TagMan probes were used. Identification of A and K alleles was performed on basis of increasing Fam and Cy5 fluorescence, respectively. The developed method was validated with PCR- RFLP analysis of 50 DNA samples from holsteinized Black-and-White cows. The developed real-time PCR-base method requires less time (up to 1 hour) in comparison with PCR-RFLP analysis and could be used for rapid and effective identification of K and A allelic variants of DGAT1 in cattle.
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