中国中药杂志

2019, v.44(13) 2777-2784

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半夏钙调蛋白基因的克隆与表达分析
Cloning and expression analysis of calmodulin gene from Pinellia ternate

刘兴;代欢欢;山雨思;杨怡;廖志华;吴能表;
LIU Xing;DAI Huan-huan;SHAN Yu-si;YANG Yi;LIAO Zhi-hua;WU Neng-biao;Key Laboratory of Eco-environments in Three Gorges Reservoir Region,Ministry of Education,School of Life Science,Southwest University;

摘要(Abstract):

该研究根据半夏转录组数据,克隆获得2条半夏钙调蛋白基因,分别命名为Pt Ca M1和Pt Ca M2。生物信息学分析表明,Pt Ca M1,Pt Ca M2基因的完整开放阅读框均为450 bp,编码149个氨基酸,两者编码区序列一致性为80%,氨基酸序列一致性为91%,且均含有EF-hand结构域,属于Ca M家族。采用实时荧光定量PCR分析Pt Ca Ms在不同组织和不同处理中的表达模式,结果表明Pt Ca M1,Pt Ca M2基因均在块茎中表达量最高;外源Ca Cl2处理后Pt Ca Ms的表达量与对照相比显著升高,钙离子鳌合剂EGTA、钙离子通道抑制剂La Cl3以及钙调蛋白拮抗剂TFP处理后其表达量逐渐降低。该研究从半夏中成功克隆得到Pt Ca Ms基因,为深入探索Pt Ca Ms基因功能特点和半夏生物碱合成的研究奠定基础。
According to the data of Pinellia ternate transcriptome,two calmodulin genes were cloned and named as Pt Ca M1 and PtCa M2 respectively. The results of bioinformatics analysis showed that Pt Ca Ms genes contained a 450 bp open reading frame,encoding149 amino acids.The identity of the coding sequences was 80%,and the identity of amino acids sequence was 91%. Pt Ca Ms genes contained EF-hand structure domain,belonging to the Ca M families. The Real-time PCR analysed the expression patterns of Pt Ca Ms in different tissues and different treatments. Results showed that Pt Ca M1 and Pt Ca M2 gene were the highest expression level in tuber. Under Ca Cl2 treatment,the expressions of Pt Ca Ms were significantly higher than the control. Under EGTA,La Cl3 and TFP treatments,the expression level of Pt Ca Ms decreased gradually. In this study,the Pt Ca Ms gene were successfully cloned from P. ternate,which laid a foundation for the functional characteristic of Pt Ca Ms gene and the synthesis of alkaloids from P. ternata for further study.

关键词(KeyWords): 半夏;钙调蛋白;基因克隆;表达分析
Pinellia ternate;calmodulin;cloning;gene expreesion

Abstract:

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基金项目(Foundation): 国家自然科学基金项目(30500041)

作者(Author): 刘兴;代欢欢;山雨思;杨怡;廖志华;吴能表;
LIU Xing;DAI Huan-huan;SHAN Yu-si;YANG Yi;LIAO Zhi-hua;WU Neng-biao;Key Laboratory of Eco-environments in Three Gorges Reservoir Region,Ministry of Education,School of Life Science,Southwest University;

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