Simultaneous genotyping of 4 SNPs in promoter III of the ovine ACACA

• Published in: Small ruminant research Vol. 138; pp. 25 - 30
• Main Authors: Sztankoova, Z., Rychtarova, J., Kyselová, J., Czerneková, V.
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.05.2016
• Subjects: Acetyl Co-A carboxylase; PEA; Polymorphism; Sheep; Sheep; PEA; Acetyl Co-A carboxylase; Polymorphism
• ISSN: 0921-4488; 1879-0941
• DOI: 10.1016/j.smallrumres.2016.03.027

•The genetic variability in the PIII promotor of the ACACA gene in different sheep breeds kept in the Czech Republic.•By sequencing analysis has been identified 2 novel SNPs.•Developed rapid, efficient and sensitive method for typing 4 SNPs using PEA.•PEA will serve as a useful genotyping technique for routine simultaneous typing of 4 genetic variants in a single gene.•Genotype AG/GT/CG/CC occurred in all 4 observed sheep breeds. The acetyl-CoA carboxylase-α (ACACA) gene is the rate-limiting enzyme for the biosynthesis of fatty acids in animal tissues. By sequencing analysis in promoter III of the ovine ACACA we identified 2 novel SNPs (A>G at 1168bp and a T nucleotide insertion at 1182bp) (GenBank accession number AJ292286). The novel variant (A>G at 1168bp) was present in Cigaja, Sumavka, and Improved Valachian breed kept in the Czech Republic and T insertion at 1182bp was present in the East Friesian breed. A protocol for rapid efficient simultaneous genotyping of ACACA variants using the primer extension method (PEA) was described, and a total of 171 animals belonging to sheep breeds: Cigaja (9), Sumavka (87), East Friesian (59) and Improved Valachian (16) were genotyped. Variant alleles are: A/G at 1168bp, G/T at 1330bp, C/G at 1138bp and C/T at 1430bp.The genotype combination AG/GT/CG/CC occurred in all 4 breeds kept in the Czech Republic with frequency of occurrence ranging from 15.25% to 44.45%. Allele frequencies of the Cigaja breed differ significantly from the other Czech sheep breeds.