DETERMINATION OF MITOCHONDRIAL CYTOCHROME-B GENE SEQUENCE FOR RED DEER (CERVUS-ELAPHUS) AND THE DIFFERENTIATION OF CLOSELY-RELATED DEER MEATS

The cytochrome b gene sequence for red deer was determined using the D ye Terminator Cycle Sequencing method and used for identification of d eer 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, p igs, sheep and goats, respectively. To differentiate the deer meat, ol igonucleotide primers RD-1 (5'-TCATCGCAGCACTCGCTATAGTACACT-3'), RD-2(5 '-ATCTCCAAGTAGGTCTGGTGCGAATAA-3') were designed for the region of the cytochrome b gene of red neer. 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 PC R products, red deer could still be detected in meat heated at 120 deg rees C. To discriminate between red and sika deer, these PCR products were digested by a restriction enzyme (EcoRI,BamHI,ScaI) and analyzed by 4% agarose gel electrophoresis. As a result, the red deer fragment was digested by EcoRI to 67/127 bp fragments but not by BamHI and Seal . The sika deer fragment was digested to 48/146 bp and 49/145 bp fragm ents with the two other enzymes, and thus it is possible to differenti ate between the two kinds of deer from the digestion pattern of restri ction enzymes, (C) 1998 Elsevier Science Ltd. All rights reserved.