Identification Of Animal Materials In Traditional Chinese Medicine Based On DNA Barcoding Technology

objective: To discuss the feasibility of using mtCO I gene sequence toefficiently identify antelope horn and scorpio in Traditional Chinesemedicine preparation.Methods: The population identification barcodes were constructed invaried Samples of Antelope horn and Scorpion from different producers, throughadequately grinding, extracting the genome DNA by DNA extraction kit, andbased on C0I nucleotide sequences by PCR amplification with the primer LCO1490and HC02198or0703. The PCR products were sequenced directly. BLASTcomparison was made between the sequencing results and the DNA barcodes ingenebank. The phylogenetic trees were constructed with the same tree (DNAMAN)and Neighbor-joining(NJ, MEGA5.2) method.The sequences of Saiga tatarica andScorpionidae from NCBI were selected to compare with standard medicinalmaterials of Antelope horn and Scorpion, in order to checkout the Authenticityof Antelope horn and Scorpionidae in Traditional Chinese medicine preparation.And the genetic distance was calculated using the Kimura-2-parameter.Results: Original nucleotide sequence data of partial mitochondrial CO1gene，about658base pairs, were collected from8Traditional Chinese medicinepreparation samples of Antelope horn. The phylogenetic tree showed that theSamples of Traditional Chinese Medicine preparation and reference substance assembled at first，and then assembled with the outgroup. The intraspecificgenetic distances of these samples ranging from0～0.064with an average of0.02, and the interspecific genetic distances ranging from0.167～0.195.575base pairs of partial mtCO I gene were collected from6Traditional ChineseMedicine preparation samples of Scorpion. The phylogenetic tree showed thatthe Samples of Traditional Chinese Medicine preparation and referencesubstance assembled at first，and then assembled with the outgroup. Theintraspecific genetic distances of these samples ranging from0～0.021withan average of0.02, and the interspecific genetic distances ranging from0.199～0.207.Conclusion: The results of our research suggested that mtCO I gene wasa valid DNA barcoding gene for Antelope horn and Scorpion identification inTraditional Chinese Medicine preparation. Because the same genes could beobtained in other organs of antelope, other detection methods could be appliedsimultaneously to ensure the accuracy of the results.