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[红花生育三烯酚环化酶基因的克隆与表达分析]

[Cloning and expression analysis of tocopherol cyclase gene in Carthamus tinctorius].

作者信息

Guan Li-Li, Cui Qi, Han Yu-Tong, Wu Yun-Yun, Hu Ren-Ge, Gu Tian-Yao, Li Hai-Yan, Li Xiao-Kun

机构信息

College of Life Science, Jilin Agricultural University, Changchun 130118, China.

High School Attached to Northeast Normal University Experimental, Changchun 130118, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2016 Oct;41(20):3727-3732. doi: 10.4268/cjcmm20162005.

Abstract

The tocopherol cyclase was one of the key enzymes in plant vitamin E biosynthesis pathway. According to the study of Carthamus tinctorius transcriptome data,the Tocopherol cyclase gene was obtained using RT-PCR techniques and named CtTC . Bioinformatics analysis showed theopen reading frame (ORF)of CtTC was 1 524 bp. The putative protein contained 507 amino acids with a predicted molecular mass of 62.9 kDa and theoretically isoelectric point was 5.01.Signal peptide analysis showed that it was a non secretory protein, and there was no signal peptide. The subcellular localization showed that the CtTC protein was located in the chloroplast. The expression of CtTC gene in safflower seeds at different development stages was determined by quantitative real-time PCR, it was found that the highest expression level of CtTC gene was detected in 50 DAF.Quantitative RT-PCR analysis suggested that expression of CtTC is induced and strengthened by drought stresses. This research provided a candidate gene for metabolic engineering of vitamin E and resisting stress.

摘要

生育三烯酚环化酶是植物维生素E生物合成途径中的关键酶之一。根据对红花转录组数据的研究,利用RT-PCR技术获得了生育三烯酚环化酶基因,并将其命名为CtTC。生物信息学分析表明,CtTC的开放阅读框(ORF)为1524 bp。推测的蛋白质含有507个氨基酸,预测分子量为62.9 kDa,理论等电点为5.01。信号肽分析表明它是一种非分泌蛋白,不存在信号肽。亚细胞定位表明,CtTC蛋白位于叶绿体中。通过实时定量PCR测定了CtTC基因在红花种子不同发育阶段的表达,发现CtTC基因在开花后50天表达水平最高。定量RT-PCR分析表明,干旱胁迫可诱导并增强CtTC的表达。本研究为维生素E代谢工程和抗逆性提供了一个候选基因。

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