Molecular Phylogenies Study Of Rana Animals And DNA Fingerprinting Application In The Discrimination Of Oviductus Ranae

As a valuable Traditional Chinese Medicines, Oviductus Ranae—oviduct of female Rana is very expensive, which is the world's only valuable economic frog species that can be used as medicine, health care products and nourishment. As an important economic species, the Changbai subspecies of Oviductus Ranae are mainly distributed in the Changbai Mountain area in Northeast China. Oviductus Ranae has been used for thousands of years and is still being widely used at present.There are two kinds of Oviductus Ranae modalities on the market:original drying animal and fat peeled (female oviduct). Similar products, such as Rana amurensis, fake Rana nigromaculata and Buffo bufo gargarizans are on the market at present. Identification on these medicinal materials is still mainly based on shape features, microscopic characteristics, and physicochemical properties. But because of the lack of obvious morphological identification features and special features chemical compositions, the identification of Oviductus Ranae is still very difficult in practice.On the basis of animal genomes totipotency, rana genome was extracted from somatic cells and oviducts cells as template DNA for PCR.30 random primers with good polymorphism are screened out from 150 random primers. According to the RAPD amplifications of 19 samples, we picked out 8 results for polymorphism analysis.8 random primers were used in the amplification of 19 samples, which obtained 786 polymorphic amplified fragments that contained 120 different bands.Samples No.1,2,3 are genomes of Changbai subspecies. A specific band of 2100bp was able to be amplified by random primer S131, which could be used as potential gene target for the distinction of Chanbai subspecies and other samples. Meanwhile, primer S131 is also a potential identify primer for Chanbai subspecies. There is another specific band of 2000bp of sample No.16 amplified by random primer S339. Consequently, S339 may be an identify primer to distinguish Rana catesbiana from others. A Rapid Guide is summarized by all these specific bands. When a sample needs to be identified, we can extract its genome and conduct RAPD amplification by these 8 random primers. Therefore the results could be judged by the corresponding bands and this rapid guide.In order to study the phylogenetic status and species effectiveness of Rana, we investigated the phylogeny of 18S rRNA of 14 samples by Maximum Parsimony, Maximum Likelihood and Neighbor-joining analyses. The results suggested the species group division of rana chensinensis should be re-evaluated within a phylogenetic context. We were able to get the specific DNA information easily, hence to do the species identification and phylogenetic studies.