摘要： 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1 014 bp, 编码338个氨基酸,推测蛋白分子量为38.46 kDa,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(100,300,500 mmol/L)和不同时间UV(12,24,36 h)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。

关键词: 长叶红砂, F3H基因, 定量RT-PCR, 真核表达载体

Abstract: Reaumuria trigyna is a small shrub endemic to the Eastern Alxa-Western Ordos area in Inner Mongolia. R.trigyna has a vital ecological function due to its remarkable capability of salt and drought tolerance.Additionally, R.trigyna had been used as traditional herb for the treatment of eczema,dermatitis and other diseases.Flavanone-3-hydroxylase(F3H)is an essential and key enzyme in the flavonoid biosynthetic pathway,it catalyzes the 3-hydroxylation of flavanones.An open reading frame(ORF)of RtF3H3 has been isolated which encodes a protein with 338 amino acids.The theoretical isoelectric point(pI)of RtF3H3 was predicted to be 5.7 and the putative molecular weights was predicted to be 38.46 kDa.The analysis of expressions level of RtF3H3 by quantitative RT-PCR showed that the high transcript levels in stems but low transcript levels in roots and leaves,and the expression level of RtF3H3 was induced by NaCl concentrations and ultraviolet(UV)treatments.In additional,constructing yeast expression vectors of RtF3H3 to lay a foundation for further study of RtF3H3.

Key words: Reaumuria trigyna, F3H gene, Quantitative RT-PCR, Expression vector

中图分类号: 

• Q943.2