Isolation Of Genes Related To Dragon Biosynthesis From Dracaena Cambodiana Pierre Ex Gagnep

Dracaena draco Linn. is the peculiarly rare plant of tropical and subtropical area. It is the important material of famous civilian medicament dragon's blood. The dragon's blood plays an important role in the Chinese medicinal materials market. At present, the dragon's blood is extracted from the stem with lipid of perennial Dracaena draco Linn., and the method needs many raw materials. The Dracaena draco Linn. is faced with dried up menace because of continuing lopping, and it has been to the endangered plant under third grade state protection. How to effective exploit and utilize the resource of Dracaena draco Linn., and to be continuable and high effective using are the important aspect of research. To isolate and identify the genes related to dragon's blood biosynthesis from Dracaena draco Linn. will help to reveal the mechanism of dragon's blood forming,and to offer theory base for increasing the content of dragon's blood and inventing the new dragon's blood production mode.The study is based on the materials which are induced by inducer and non-induced from the Dracaena cambodiana seedlings in tissue culture. The study is initiated by construction a subtraction cDNA library of genes related to dragon's blood biosynthesis from Dracaena draco Linn. by using suppressive subtractive hybridization (SSH) methodology, then 431 recombinants are got by screening the subtraction cDNA library with PCR. To randomly pick the 200 clones by Reverse Northern Blot, and to eliminate the pseudo positive, then 59 positive clones of obvious difference are got, while after sequencing, 51 different fragments are got. To comparatively analyze the different fragments by BLASTn and BLASTx, the result shows that: 5 cDNA fragments related to signal transduction, 5 cDNA fragments related to transcription and translation protein, 12 cDNA fragments related to substance and energy metabolism enzyme, and other 23 cDNA fragments which can not be found from the Genbank with the significantly related homologous sequence. To speculate that they may be the new genes of unknown function or gene fragments nearing 3'-end, and the homology is very low, so it is difficult to speculate the function by homology comparison.