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长叶红砂黄烷酮-3-羟基转移酶3基因(RtF3H3)的克隆、表达分析及酵母表达载体的构建

DOI: 10.7668/hbnxb.2015.05.008, PP. 45-50

Keywords: 长叶红砂,F3H基因,定量RT-PCR,真核表达载体

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Abstract:

黄烷酮-3-羟基转移酶(F3H)是植物黄酮合成途径上游的一个关键酶,它催化黄烷酮C3位羟化,形成二氢黄酮醇。构建长叶红砂RtF3H3基因酵母表达载体,为进一步研究RtF3H3基因功能奠定基础。基于高通量转录组测序结果,利用RT-PCR技术克隆获得长叶红砂F3H3(RtF3H3,GenBank登录号LC055546)基因,其开放阅读框长1014bp,编码338个氨基酸,推测蛋白分子量为38.46kDa,理论等电点为5.7。通过定量RT-PCR研究发现,RtF3H3在茎中表达量较高,在根和叶中表达量较低。不同程度NaCl(100,300,500mmol/L)和不同时间UV(12,24,36h)胁迫都能诱导RtF3H3基因表达量升高。另外,构建了RtF3H3基因酵母表达载体,为进一步研究RtF3H3基因功能奠定基础。

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