Determination of mitochondrial cytochrome B gene sequence for red deer ( Cervus elaphus) and the differentiation of closely related deer meats

Abstract

The cytochrome b gene sequence for red deer was determined using the Dye Terminator Cycle Sequencing method and used for identification of deer meat in meat and meat products. Red deer showed a similarity of 94.1, 84.0, 81.1, 85.5 and 85.6% to sika deer ( Cervus nippon), bovine, pigs, sheep and goats, respectively. To differentiate the deer meat, oligonucleotide primers RD-1(5′-TCATCGCAGCACTCGCTATAGTACACT-3′), RD-2(5′-ATCTCCAAGTAGGTCTGGTGCGAATAA-3′) were designed for the region of the cytochrome b gene of red deer. The PCR amplified 194 bp fragments from red and sika deer, but no fragments from bovine, pig, chicken, sheep, goat, horse and rabbit DNA. Although cooking the meats reduced the PCR products, red deer could still be detected in meat heated at 120 °C. To discriminate between red and sika deer, these PCR products were digested by a restriction enzyme ( EcoRI, BamHI, ScaI) and analyzed by 4% agorose gel electrophoresis. As a result, the red deer fragment was digested by EcoRI to 67/127 bp fragments but not by BamHI and ScaI. The sika deer fragment was digested to 48/146 bp and 49/145 bp fragments with the two other enzymes, and thus it is possible to differentiate between the two kinds of deer from the digestion pattern of restriction enzymes.