唐美琼, 闵丹丹, 李刚, 蒋妮, 叶云峰. 三七病程相关蛋白PR10-1基因克隆及功能初步分析J. 药学学报, 2015,50(2): 227-232.
引用本文: 唐美琼, 闵丹丹, 李刚, 蒋妮, 叶云峰. 三七病程相关蛋白PR10-1基因克隆及功能初步分析J. 药学学报, 2015,50(2): 227-232.
TANG Mei-qiong, MIN Dan-dan, LI Gang, JIANG Ni, YE Yun-feng. Cloning and functional characterization of pathogenesis-related PR10-1 gene in Panax notoginsengJ. Acta Pharmaceutica Sinica, 2015,50(2): 227-232.
Citation: TANG Mei-qiong, MIN Dan-dan, LI Gang, JIANG Ni, YE Yun-feng. Cloning and functional characterization of pathogenesis-related PR10-1 gene in Panax notoginsengJ. Acta Pharmaceutica Sinica, 2015,50(2): 227-232.

三七病程相关蛋白PR10-1基因克隆及功能初步分析

Cloning and functional characterization of pathogenesis-related PR10-1 gene in Panax notoginseng

  • 摘要: 采用同源克隆法和RACE技术相结合, 对三七根部蛋白质组差异进行研究, 鉴定PR10-1蛋白相应的基因全长cDNA, 初步分析该基因在三七中的功能。测序结果显示该基因序列全长863 bp, 包含1个序列长465 bp编码155个氨基酸的开放阅读框, 命名为PnPR10-1, GenBank登录号KJ741402。 氨基酸序列同源性及系统发育树分析发现, PnPR10-1与人参的PR10-1蛋白同源性最高, 含有Bet-v-I家族等多个保守结构域。实时荧光定量PCR分析显示, PnPR10-1在1~3年生三七各组织中均有本底表达, 暗示其参与生长发育、代谢调控等生物学过程; PnPR10-1在根部受根腐病原菌(Fusarium oxysporum)胁迫上调表达, 推测PnPR10-1基因可能参与抗三七根腐病等广谱抗病防御反应。

     

    Abstract: With homology cloning approaches coupling with RACE (rapid-amplification of cDNA ends) techniques, the full-length coding sequence of pathogenesis-related protein PR10-1 with differential expression was cloned from the total RNA of the root of Panax notoginseng, and its function was explored furtherly. As a result, the longest 465 bp ORF (named as PnPR10-1 with the Accession No. KJ741402 in GenBank) was detected from the cloned sequence with full-length of cDNA of 863 bp. The corresponding peptide encoded consisted of 155 amino acids, contained some domains such as Bet-v-I, and showed high similarity with that from Panax ginseng by analysis of phylogenetic trees created from the alignments. Real-time quantitative PCR showed that the expression of PnPR10-1 gene was constitutive in different tissues of 1-3 year old plant, suggesting that it might be involved in growth, development, and secondary metabolism; yet it was up-regulated significantly with the infection of Fusarium oxysporum in root, suggesting that it might be involved in defense against many diseases including root rot in P. notoginseng.

     

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