[Cloning and bioinformatics analysis of 2-c-methyl-D-erythritol 2,4-cyclodiphosphate synthase gene in Salvia miltiorrhiza].
OBJECTIVE
To clone and analysis a 2-C-methyl-D-erythritol 2,4-cyclodiphosphate synthase (SmMCS) full-length eDNA from Salvia miltiorrhiza hairy roots.


METHODS
A full-length eDNA of SmMCS has been cloned by designing specific primers according to the transcriptome database and using the RACE strategy. ORF Finder was used to find the open reading frame of SmMCS cDNA and ClustalW has been performed to analysis the multiple amino acid sequence alignment. Phylogenetic tree has been constructed using MEGA 5. 1. Real-time quantitative PCR have been applied to detect the transcription level of SmMCS from hairy roots after elicitor Ag+ supplied.


RESULTS
The SmMCS cDNA sequence was obtained. The full length of SmMCS (DNA was 988 bp encoding 234 amino acids. The deduced protein had isoelectric point (pI) of 8.53 and a calculated molecular weight about 24. 6 kDa. Results of real time PCR indicated that elicitor of Ag+ stimulated the increase of mRNA expression of SmMCS in hairy roots, and were increased dramatically at 12 h.


CONCLUSIONS
The full-length cDNA of SmMCS was cloned from S. miltiorrhiza hairy root,which can provide a gene target for further studies of tanshinones biosynthesis and terpenoid secondary metabolites.
School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China. weigao@ccmu.edu.cn
Wei
W
Gao
Qi-Qing
QQ
Cheng
Xiao-Hui
XH
Ma
Yun-Fei
YF
He
Chao
C
Jiang
Yuan
Y
Yuan
Lu-Qi
LQ
Huang
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D019295:Computational Biology; D018506:Gene Expression Regulation, Plant; D008969:Molecular Sequence Data; D019761:Phosphorus-Oxygen Lyases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011487:Protein Conformation; D027561:Salvia miltiorrhiza / Q000737:chemistry / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D016415:Sequence Alignment
D010940:Plant Proteins; D019761:Phosphorus-Oxygen Lyases