白木香2个小分子热激蛋白基因的克隆及表达分析

发布时间：2018-06-03 04:02

  本文选题：白木香 + sHSP基因　； 参考：《中草药》2016年22期

【摘要】：目的从白木香Aquilaria sinensis中克隆sHSP1和sHSP2基因,并对它们的生物信息学及表达模式进行分析。方法根据本课题组白木香转录组数据库进行分析获得2条具有小分子热激蛋白(sHSPs)保守结构域的sHSPs基因序列,利用RT-PCR技术克隆sHSP1和sHSP2基因的全长c DNA序列,并对其进行生物信息学分析;利用实时荧光定量PCR检测sHSP1和sHSP2基因在不同组织和高盐及外源ABA、SA、MJ处理下不同时间的表达差异。结果克隆得到的白木香sHSP1和sHSP2基因开放阅读框都为474 bp,编码157个氨基酸。组织表达分析的结果显示,sHSP1和sHSP2基因主要在根中表达,在茎和叶中的表达量较少;白木香愈伤组织在高盐处理下,sHSP1和sHSP2基因分别在36 h和24 h达到最高表达水平,而愈伤组织在外源ABA、MJ处理下,sHSP1和sHSP2基因都在12 h达到最高表达水平,在SA处理下,sHSP1和sHSP2基因分别在12、24 h达到最高表达水平。结论获得了sHSP1和sHSP2 2基因全长c DNA序列,且2个基因在不同组织根、茎、叶中的表达存在差异性,在受到高盐和外源ABA、SA、MJ处理时,在不同时间的愈伤组织中的表达及积累情况也不同,该研究为后续深入研究白木香防御反应奠定了基础。
[Abstract]:Objective to clone sHSP1 and sHSP2 genes from Aquilaria sinensis and analyze their bioinformatics and expression patterns. Methods two sHSPs gene sequences with small molecular heat shock protein (HSPs) conserved domain were obtained by analyzing the transcriptional database of the white wood incense. The full-length c DNA sequences of sHSP1 and sHSP2 genes were cloned by RT-PCR technique. The expression of sHSP1 and sHSP2 genes in different tissues, high salt and exogenous ABASAMJ were detected by real-time fluorescence quantitative PCR. Results the open reading frames of sHSP1 and sHSP2 genes were 474 BP, encoding 157 amino acids. The results of tissue expression analysis showed that HSP1 and sHSP2 genes were mainly expressed in roots, but less in stems and leaves, and the highest expression levels of HSP1 and sHSP2 genes were found in callus of Aucklanthus chinensis under high salt treatment at 36h and 24h, respectively. However, the highest expression level of HSP1 and sHSP2 genes in callus was reached at 12 h after exogenous ABAA-MJ treatment, and the highest expression level of HSP1 gene and sHSP2 gene were reached at 1224 h after SA treatment. Conclusion the full-length c DNA sequences of sHSP1 and sHSP2 2 genes were obtained, and there were differences in the expression of the two genes in root, stem and leaf of different tissues. The expression and accumulation of callus were different at different time, which laid a foundation for further study on the defense response of Amur.
【作者单位】： 北京中医药大学中药学院;北京中医药大学中药现代研究中心;
【基金】：北京市自然科学基金青年项目(7154216)
【分类号】：S567.19

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