Cloning And Bioinformatic Analysis Of The Differenital Sequence From The Pistil And Scape Of Lycoris Aurea Herb

Lycoris aurea Herb. contains alkaloids, polysaccharides, flavonoids, amino acids, lectins element etc. Besides,the Galanthamine content is high ahead of other Lycoris species.It has been shown to possess dual effects such as reversible and selective inhibition of acetylcholinesterase (AChE) and adjusting the nicotinic acetylcholine receptor, and is one of the best drug to treat Alzheimer’s disease (Alzheimer’s Disease, AD).By analyzing correlative literature, we concluded that the Galanthamine content in the pistil of Lycoris aurea Herb in July was always higher than in the stem and especially highest in the scape. It tells us that there are maybe some difference in the key enzyme of galanthamine synthesis in the biosynthetic pathway and metabolic pathway in different organs in the Lycoris aurea Herb.Therefore, this thesis has applied mRNA differential display technology to display the differential sequence of the pistil and scape of Lycoris aurea Herb. Furthermore, the differential sequences were cloned and then analyzed basing on bioinformatics methods. The main results were as follow:First of all, Lycoris aurea Herb was gathered from Hengshan, then identified by Professor Pan Qingping of University of Chinese medicine. By using the RNA plant Kit, CTAB method and SDS method respectively to extract the total RNA of the pistil and scape of Lycoris aurea Herb,the RNA plant Kit method showed optimal.2. The PCR reaction conditions were optimized and then the optimum condition was determined. By Screening78pairs of primers, the differential sequences of the pistil and scape of Lycoris aurea Herb were found. Next, the differential sequences were purified and cloned.3. The positive recombinant clones were identified and sequenced.After this,the44sequencing fragments were compared by BLAST. Then we analysed the function of the proteins encoded by the the differential sequence.What is more, we had studied whether or not the differential sequence coded the related synthesis enzymes of galanthamine.