Cloning And Expression Analysis Of DCDPK2 Gene In Dracaena Draco

Dragon’s blood is a precious traditional Chinese medicine, has pharmacological functions of arresting bleeding、invigorating blood and removing blood stasis、antitumor,etc. At present, it is mainly extracted from Dracaena draco. With the increase of demand of dragon’s blood, Dracaena draco has been over-harvesting. Dracaena draco resources to be exhausted, which will reduce the production of Dragon’s Blood. Accordingly, It is an important project in present research domain to reveal the mechanisms of forming dragon’s blood which can provide theoretical foundation for increasing contents of the dragon’s blood and inventing a new manner for its production which has important theoretical and practical significance.Previous study in the laboratory, we have obtained many gene fragments related to dragon’s blood biosynthesis by constructing SSH library. From which we found that CDPK gene is possible to play an important roles on the biosynthesis of dragon’s blood. On the basis of this study, cloning full-length cDNA of CDPK gene, and for RT-PCR expression analysis,To further clarify the gene induction pathway in the Dragon’s blood to provide a theoretical basis for the role.Firstly, the design of degenerate primers to clone the gene a conservative length of 728bp fragment.Secondly,the gene 3’end of 596bp and 5’end of 615bp sequences were cloned by RACE. Finally, using RT-RCR to obtain full-length cDNA, Named DCDPK2. The gene length was 1809bp, coding region of a length of 1587bp, encoding 528 amino acids. Its encoded amino acid sequences have been reported in all the typical structural features of the CDPK genes. The protein sequence alignment was found with the CDPK genes of other plants identity is as high as 75%.Semi-quantitative RT-PCR expression analysis used Dracaena tissue culture plantlets which not been induced by treatment and after induction treatment,found the DCDPK2 gene expression markedly increased in tissue culture the addition of inducer. Then,the result of expression analysis in different processing and different parts showed that the expression of DCDPK2 gene in induced Dracaena draco stem tissue culture strdnger expression of the leaf and the roots, stems, leaves area is not induced tissue culture.