The development of methods for the isolation of DNA and identification of missense variants from hair roots of sows
2009
Hoshino, Y., Meiji Univ., Kawasaki, Kanagawa (Japan). School of Agriculture | Mizoguchi, Y. | Sasaki, Y. | Koketsu, Y.
The objectives of the present study were 1) to develop a method for the isolation of DNA from hair roots in sows fed in commercial herds, and 2) to develop a polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) method for the identification of a missense variant. We chose the porcine melanocortin-4 receptor (MC4R) gene as the first candidate to connect reproductive performance of female pigs to gene information. The MC4R gene is a key factor in the regulation of energy balance and body weight in mammals. A missense variant of the porcine MC4R gene is associated with fatness and growth in fattening pigs. Additionally, collecting hair root samples is easier than taking blood samples on commercial herds. We visited a large commercial herd raising Fsub(1) crossbreds of Landrace and Large White and Landrace sows. We collected hair roots in 3 Fsub(1) crossbred sows and 2 Landrace sows. We used 5 to 30 hair roots per sow for analysis. The MC4R region in the DNA extracted from the hair roots was amplified by PCR. The PCR product was digested with Taq I enzyme overnight at 65degC. The digested fragments were separated by electrophoresis in a 2% agarose gel. The range of amounts of DNA extracted from one hair root is between 70 and 1900 ng. Furthermore, we found the mutation of the MC4R gene in Landrace sows.
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