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Biotechnology
Year: 2018  |  Volume: 17  |  Issue: 1  |  Page No.: 26 - 34

Molecular Cloning and Expression of a Squalene Epoxidase Gene from Ilex cornuta

Li Zhu, Liangqiong Ma, Feng Xu and Weiwei Zhang    

Abstract: Background and Objective: Ilex cornuta (I. cornuta) is a medicinal plant that contains triterpenoid compounds as its pharmacologically active ingredients. Squalene epoxidase (SE) is a key enzyme in the triterpenoid biosynthesis pathway. The current study aimed to characterize a SE gene from I. cornuta. Methodology: IcSE2 was isolated from I. cornuta. It was predicted the secondary and tertiary structures of the IcSE2 protein, performed multiple sequence alignments and generated a phylogenetic tree. The expression level of IcSE2 was examined via Quantitative Real Time-PCR (qRT-PCR). Results: The cDNA sequence of IcSE2 was 2058 bp, with an open reading frame of approximately 1605 bp that codes for 534 amino acids. The predicted theoretical isoelectric point and molecular weight are 8.30 and 58.6 kDa, respectively. RxR, Flavin Adenine Dinucleotide (FAD) and Nicotinamide Adenine Dinucleotide Phosphate (NADPH) domains could be found in the deduced IcSE2 protein. Phylogenetic analysis showed that IcSE2 is closely related with the SEs from Araliaceae plants. IcSE2 expression level was the highest in roots, followed by leaves and male flowers and lowest in stems. Conclusion: The IcSE2 gene from I. cornuta was cloned and characterized for the first time. IcSE2 was strongly expressed in roots and leaves. These results may lay the foundation for studying the molecular regulatory mechanism of triterpenoid saponins in I. cornuta.

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