Prediction of CpG islands and functional verification on promoters of FPS, SS, SE in Eleutherococcus senticosus / 中草药

Objective: To obtain the distribution and functional activity of CpG islands in promoters of FPS, SS, and SE from Eleutherococcus Senticosus. Methods: Based on the promoter sequence of FPS, SS and SE, CpG islands were predicted by using EMBOSS and Li Lab. The functional verification of transformed Arabidopsis thaliana was mediated by Agrobacterium tumefaciens by using GUS and pCAMBIA1301 plasmid as the reporter gene and expression vector respectively. Results: Two CpG islands were found in FPS and SS promoter with the lengths of 520 bp, 218 bp and 108 bp, 103 bp, and three CpG islands in SE promoter in 290 bp, 119 bp and 149bp. The promoters of FPS, SS and SE all had promoter activities at different level, in which SS promoter was the highest one. Conclusion: The functional verification and distributions of CpG islands in the promoters area of FPS, SS, and SE were reported in this research at first time, which established the foundation for the methylation analysis of FPS, SS, and SE and the further studying of the mechanism expression regulation in E. Senticosus.

Analysis on structure and phylogeny of chloroplast genomes in Araliaceae species / 中草药

Objective: To obtain the structural characteristics and phylogentic relationships of the chloroplast genome in Araliaceae species. Methods: we used 20 chloroplast genomes which have sequenced as materials, they were from 20 species belonging to 10 genus of Araliaceae. Analysis the differences of genomes and the dilation or shrink of four boundaries for IR, we used MEGA 4.0 to build the phylogenetic tree with Angelica gigas of sibling species as the outgroup and analysis their phylogentic relationships. Results: There was a small difference among the chloroplast genomes size, and the largest difference is 1 909 bp. All of species had existed gene replacements, cemA replaced ycf10, and number of genes existed some differences, they were mainly caused by tRNA. The four boundaries of IR was relatively conservative, only Panax vietnamensi, Panax notoginseng and Schefflera delavayi were special, their boundary genes were in IR. All nodes of the phylogenetic tree of Araliaceae which was based on Angelica gigas were of high supports, and the tree had good resolution to reflect the genetic relationships among Araliaceae. Conclusion: Chloroplast genomes have a lot of information, it can be used to analysis phylogeny among the species which are affinity or faster evolution.

Cloning and bioinformatic analysis of mevalonate diphosphate decarboxylase gene promoter in Eleutherococcus senticosus / 中草药

Objective: To clone and analyze the sequence of mevalonate diphosphate decarboxylase (MDD) gene promoter from Eleutherococcus senticosus. Methods: According to the full length of MDD cDNA sequence, using PCR cloning and TAIL-PCR methods, we are cloning 5'DNA sequence and promoter sequence of MDD gene. And bioinformatic analysis is used. Such as PlantCARE. Results: We have cloned the 5'DNA sequence successfully, with full length of 1024 bp. And the promoter sequence is 1423 bp. Bioinformatic analysis shows that the promoter sequence has 49 TATA-box and 25 CAAT-box. In addition, the promoter has some cis-acting elements responsive to abscisic acid, environmental stresses, MeJA, required for endosperm expression, light-regulated and so on. Last but not least, there have two MYBHv1 and two MYB binding sites. Conclusion: We have firstly cloned and analyzed the E. senticosus MDD gene promoter sequence. It is important to it's expression.