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植物抗病性信号传导调控基因的克隆与表达检测方法的研究

DOI: 10.7685/j.issn.1000-2030.2003.04.008, PP. 30-35

Keywords: 植物抗病性信号传导,调控基因,克隆,表达谱,半定量RT-PCR

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Abstract:

建立了从不同植物中克隆抗病性信号传导调控基因及检测其表达的方法,并优化了相关条件。用RTPCR和PCR方法从拟南芥、烟草、番茄、中华鳖中克隆了7个信号传导通路中18个调控基因的cDNA和DNA序列,它们分别是抗病基因Pto介导的蛋白激酶级联中的Pti4、Pti5和Pti6,细胞编程死亡通路中的Hin1和hsr203,水杨酸介导的系统性获得抗病性通路中的NPR1,乙烯信号通路中的ETR1、ERS1、CTR1、EIN2和ERF1,茉莉酸信号通路中的COI1,核黄素信号通路中的RFBP和LR,脱落酸信号通路中的ABF3、ABF4和ABH1,以及调控不同信号通路的通调基因NDR1。进一步研究了这些基因表达的定量测定方法用细菌激发子harpinEa处理植物,提取RNA作为cDNA第1链合成的模板,以在真核生物中广泛同源和高度保守的细胞伸长翻译因子基因EF1α为内参,用半定量RTPCR法测定基因mRNA的积累,可以比较准确地检测不同基因的相对表达水平。

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