Studies On Identification Of Traditional Chinese Medicine Snake Gallbladder, Deer By Molecular Marker And Molecular Systematics Of Cervinae

Snake gallbladder is a traditional Chinese medicine. The researches on chemical components, pharmacological effect and authentication of this Chinese crude drug were reviewed. DNA fragment of 12S rRNA gene was amplified from the templates extracted from the membrane or bile of snake gallbladder, and sequenced subsequently. The sequence analysis indicates that the technique of DNA molecular marker could be used for the authentication of both snake gallbladder and bile, and it also infers the technique is adaptable for the identification of other animal secretion.In the study of the Chinese medicine made of deer, the researches on chemical constituents, pharmacological activity as well as authentication for the crude drugs were also summarized. To develop a convenient and accurate method for the identification of the crude drugs originated from deer, including pilose antler, penis and testis, tendon and foetus. Based on the analysis of DNA sequences of mitochondrial Cyt b gene from original animals of genuine crude drugs, Cervus nippon or Cervus elaphus, and their adulterants, a pair of allele-specific primers were designed for distinguishing the genuine crude drugs by PCR method. The diagnostic PCR test for original animals and medicinal materials shows that the primers designed in present study are highly specific for Cervus nippon and Cervus elaphus, and they could be used for the authentication of traditional Chinese medicines of deer. It also demonstrates that the origins of commercial deer crude drugs are complicated and more efficient quality control is necessary for supervising the crude drug in the market.Finally, the phylogenetic relationships of four genera of subfamily Cervinae, 5species of Cervus and 6 subspecies of C. elaphus distributing in China were studied bysequence analysis of Cyt b gene. The results reveal that: 1. Axis is not monophyletic and it is strongly supported that A. porcinus should be merged into Cervus. The close genetic relationship between Elaphurus and Cervus suggests that two taxa should be incorporated same genus, whereas the classification status of Dama dama still remains uncertain. 2. The species of Cervus distributing in China make a monophyletic clade. 3. Subspecies of C. elaphus in China, deriving from C. nippon, form a monophyletic group.