Cloning and bioinformatics analysis of ent-kaurene oxidase synthase gene in Salvia miltiorrhiza.
- Author:
Ya-ting HU
;
Wei GAO
;
Yu-jia LIU
;
Qi-qing CHENG
;
Ping SU
;
Yu-zhong LIU
;
Min CHEN
- Publication Type:Journal Article
- MeSH:
Amino Acid Sequence;
Cloning, Molecular;
Computational Biology;
methods;
Cytochrome P-450 Enzyme System;
chemistry;
genetics;
Models, Molecular;
Molecular Sequence Data;
Phylogeny;
Protein Structure, Tertiary;
Salvia miltiorrhiza;
enzymology
- From:
China Journal of Chinese Materia Medica
2014;39(21):4174-4179
- CountryChina
- Language:Chinese
-
Abstract:
Based on the transcriptome database of Salvia miltiorrhiza, specific primers were designed to clone a full-length cDNA of ent-kaurene oxidase synthase (SmKOL) using the RACE strategy. ORF Finder was used to find the open reading frame of SmKOL cDNA, and ClustalW has been performed to analysis the multiple amino acid sequence alignment. Phylogenetic tree has been constructed using MEGA 5.1. The transcription level of SmKOL from the hairy roots induced by elicitor methyl jasmonate (MeJA) was qualifiedby real-time quantitative PCR. The full length of SmKOL cDNA was of 1 884 bp nucleotides encoding 519 amino acids. The molecular weight of the SmKOL protein was about 58.88 kDa with isoelectric point (pI) of 7.62. Results of real-time quantitative PCR analyses indicated that the level of SmKOL mRNA expression in hairy roots was increased by elicitor oMeJA, and reached maximum in 36 h. The full-length cDNA of SmKOL was cloned from S. miltiorrhiza hairy root, which provides a target gene for further studies of its function, gibberellin biosynthesis and regulation of secondary metabolites.
