地黄RgDXR基因的克隆及表达分析

发布时间：2018-04-25 17:20

本文选题：地黄 + RgDXR基因　；参考：《植物生理学报》2017年04期

【摘要】：本研究采用RT-PCR法对地黄DXR基因进行克隆,并用生物信息学方法分析其功能结构域、系统进化等;将地黄DXR基因克隆到原核表达载体p ET-32a上,使用大肠杆菌BL21菌株进行Rg DXR蛋白的原核表达及纯化,实时荧光定量PCR方法分析Rg DXR在不同组织、不同内生菌诱导下的表达特性。获得的Rg DXR基因c DNA全长为1 425 bp,编码474个氨基酸。生物信息学分析发现Rg DXR基因编码的蛋白质具有典型的DXR功能结构域,属于植物DXR基因家族。系统进化树分析表明,地黄Rg DXR与芝麻亲缘关系最近。荧光定量PCR结果显示,栽培品种‘北京3号’根中Rg DXR表达量最高。内生真菌诱导实验表明,Rg DXR基因的转录水平与梓醇等环烯醚萜类物质的含量成正相关。本研究克隆所得的地黄Rg DXR基因属于DXR基因家族,推测其可能与地黄中环烯醚萜类物质的生物合成有关。
[Abstract]:In this study, the DXR gene of Rehmannia glutinosa was cloned by RT-PCR, its functional domain and phylogeny were analyzed by bioinformatics, and the DXR gene of Rehmannia glutinosa was cloned into the prokaryotic expression vector p ET-32a. E. coli BL21 strain was used for prokaryotic expression and purification of rg DXR protein. The expression characteristics of rg DXR in different tissues and endophytic bacteria were analyzed by real-time fluorescence quantitative PCR. The total length of c DNA of rg DXR gene was 1 425 BP, encoding 474 amino acids. Bioinformatics analysis showed that the protein encoded by Rg DXR gene had a typical DXR functional domain and belonged to the DXR gene family of plants. Phylogenetic tree analysis showed that Rg DXR of Rehmannia glutinosa had the closest relationship with sesame seeds. The results of fluorescence quantitative PCR showed that the rg DXR expression was the highest in the roots of the cultivar 'Beijing 3'. The induction of endophytic fungi showed that the transcription level of Rg DXR gene was positively correlated with the content of iridoterpenes such as catalpol. In this study, the Rg DXR gene of Rehmannia glutinosa belongs to the DXR gene family, which may be related to the biosynthesis of the terpenoids from Rehmannia glutinosa.
【作者单位】：河南中医药大学药学院呼吸疾病诊疗与新药研发河南省协同创新中心;
【基金】：国家自然科学基金(81603232) 国家“十二五”科技支撑计划(2011BAl06802) 河南中医学院博士科研基金(BSJJ2011-07)~~
【分类号】：Q943.2;S567.234

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