Comparison And Homological Analysis On DNA Fragments Cloned Of The Radices Saussureae And Radix Linderae

As the habitat of the traditional Chinese medicine is complex, the radix saussureae lappae, aplotaxis anriculata, vladimiria denticulate ling, longpeduncle kadsura root and radix aristolochiae may be used as costustoot in the traditional Chinese medicine.Some traditional Chinese medicines are easy to be confused, such as the lindera strychnifolia and the Cocculus Laurifolius DC. It has been making the market confused and security problems happened. Therefore, the authentication of chinese medicines plays an important role in the traditional Chinese medicine domain. It is hard to control the sorts of the traditional Chinese medicine and identify the traditional Chinese medicine by physical and chemical methods. This lays down the hidden danger for the opportunistic practice. This project provided a new method on the traditional Chinese medicine by molecular identification, as basic research material for the authentication of Chinese medicines advancing to the new stage.The project's main research contents and results are as follows:1. To select the decoction pieces and fresh materials of the radices saussureae and radix linderae. They are identified by the professor Zhou Ri bao that the decoction pieces and fresh materials of the radices saussureae are all belonged to the sort of the Saussurea lappa C.B Clarke and the decoction pieces and fresh materials of the radix linderae belonged to the sort of the Lindera aggregate(Sims) Kosterm. The DNA of the decoction pieces and fresh materials both above extracted by the method of CTAB.2. To optimize the conditions of the PCR and find the best conditions for PCR, screen the random primers to find the one that can be used as DNA amplification for both the decoction pieces and fresh materials, compare with the DNA bands ampliciated to find the consensus band both.3. To recover the consensus bands from agarose gel as templates for the next step of the cloning.5. To clone the consensus bands acquired by amplifing the genomic DNA of decoction pieces and fresh materials to select the positive colony for sequencing after transformation and homological analysis by the biological information sciences. The results indicated that the similarity of sequences of radix saussureae lappae decoction pieces and fresh materials, lindera strychnifolia decoction pieces and fresh materials achieves 98% and 97% respectively. The consequences coincide with the anticipated result.The research conclusions are as follows:1. There are many differences between the DNA of the decoction pieces and fresh materials. The case of the missing of bases in decoction pieces sequences consistents with DNA degradation and incompletion after hammering.2. The results indicated that the similarity of sequences of the decoction pieces and fresh materials of the radices saussureae and radix linderae achieve above 97%. they have homologies.3. This project can slove fundamentally the prombles on the traditional Chinese medicine by molecular identification. The results are accurater and more scientific.