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北方园艺  2014 

肉桂酸-4-羟化酶基因及其启动子克隆与表达分析

, PP. 122-127

Keywords: 肉桂酸-4-羟化酶,启动子,实时荧光定量PCR

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Abstract:

以不同发育时期的果实为试材,采用两步法逆转录与抑制PCR技术相结合的方法构建均一化全长cDNA文库,筛选出一条编码肉桂酸-4-羟化酶的全长cDNA(命名为PsC4H);采用APA-Walking技术,分离获得该基因的启动子序列;采用实时荧光定量PCR技术,检测该基因在果实不同发育时期的表达动态。结果表明PsC4H基因全长为1772bp,ORF(OpenReadingFrame)为1515bp,编码504个氨基酸,相对分子量为145719.6,等电点为4.94,经序列同源性分析发现,PsC4H氨基酸序列与李属果树高度同源;经PlantCare软件预测,PsC4H基因的启动子除具有TATA/CAAT-box外,还含有G-box、HSE等特异作用元件;实时荧光定量结果显示,PsC4H在果实整个生长发育过程中呈下调-上调的表达趋势,其中在成熟果中表达量最高。

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