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Frontiers of Agriculture in China

ISSN 1673-7334

ISSN 1673-744X(Online)

CN 11-5729/S

Front Agric Chin    2011, Vol. 5 Issue (3) : 400-406     DOI: 10.1007/s11703-011-1121-3
RESEARCH ARTICLE |
Molecular cloning and characterization of GuHMGR, an HMG-CoA reductase gene from liquorice (Glycyrrhiza uralensis)
Chunying MA1, Chunsheng LIU2,3, Wenquan WANG2,3()
1. Key Laboratory of Regulation and Control of Crop Growth of Hebei Province, College of Agronomy, Agricultural University of Hebei, Baoding 071000, China; 2. School of Chinese Pharmacy, Beijing University of Chinese Medicine, Beijing 100102, China; 3. The Engineering Research Center for Chinese Medicine Standardized Production of Educational Ministry, Beijing 100102, China
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Abstract  

A full length cDNA encoding HMGR (designated as GuHMGR) was isolated from liquorice (Glycyrrhiza uralensis) based on degenerated PCR and genome walking. The full length cDNA of GuHMGR was 2330 bp with a 1518-bp open reading frame (ORF) encoding a 505-aa polypeptide. Bioinformatics analysis indicated that there were two trans-membrane domains in GuHMGR. A molecular model of tertiary structure showed that GuHMGR is a novel HMGR with a similar spatial structure to other plant HMGRs. The deduced polypeptide of GuHMGR has an isoelectric point (pI) of 6.41 and a calculated molecular weight of about 54.7 kDa. Sequence comparison and phylogenetic tree analysis showed that GuHMGR had the highest homology with HMGRs from Pisum sativum and Medicago truncatula, indicating that GuHMGR belongs to the plant HMGR group. Expression analysis showed the similar amount of transcript level of GuHMGR in roots and leaves, suggesting that this gene was expressed constitutively in plants. Therefore, this novel HMGR gene would possibly provide a new strategy for studying the glycyrrhizin metabolism at the molecular level in the future.

Keywords cloning      liquorice      glycyrrhiza uralensis      genome walker      HMG-CoA reductase     
Corresponding Authors: WANG Wenquan,Email:bushbucker@126.com; wwq57@126.com   
Issue Date: 05 September 2011
URL:  
http://academic.hep.com.cn/fag/EN/10.1007/s11703-011-1121-3     OR     http://academic.hep.com.cn/fag/EN/Y2011/V5/I3/400
Fig.1  The glycyrrhizin biosynthesis pathway. HMG-CoA stands for 3-hydroxy-3-methylglutaryl coenzyme A, MVA for mevalonic acid, IPP for isopentenyl pyrophosphate, DMAPP for dimethylallyl pyrophosphate, GPP for geranyl pyrophosphate, and FPP for farnesyl pyrophosphate, respectively. HMGR, the enzyme that catalyze the second biochemical reaction from HMG-CoA to MVA, represents 3-hydroxy-3-methylglutaryl-CoA reductase.
Fig.2  The PCR amplified pattern of the gene from Fisch. A is 2633 bp conservative sequence of the gene using the primer pair (GuHMGRF1, GuHMGRR1); B is Genome walking products of the gene. M is Molecular weight marker DL2000; 1-4 represent 2633 bp conservative sequence of the gene, the first round PCR product of genome walking, the second round PCR product of genome walking, and the third round PCR product of genome walking respectively.
Fig.3  The full-length cDNA sequence and deduced amino acid sequence of . 3-hydroxy-3-methylglutary1-CoA reductase (). The start codon (ATG) and the stop codon (TAA) are boxed.
Fig.4  Relationships between the primary amino acid sequences of HMGRs in plants.
Fig.5  The 3-D model of .
Fig.6  Expression profile of gene in new leaf, old leaf, taproot and lateral root.
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