艾纳香葡萄糖基转移酶基因的克隆与生物信息学分析

Cloning and Bioinformatics Analysis of UFGT in Blumea balsamifera

艾纳香(Blumea balsamifera)作为贵州道地药材,其次生代谢产物,如类黄酮、艾纳香素等具有重要的药理作用。葡萄糖基转移酶(UFGT)是艾纳香类黄酮代谢途径中的一个关键酶。本研究通过对艾纳香转录组进行测序,借助引物PCR成功克隆得到其葡萄糖基转移酶基因的cDNA序列,并对其进行了相应的生物信息学分析。分析发现,艾纳香UFGT基因cDNA全长1 527 bp,共编码508个氨基酸,所编码蛋白的分子量为56.155 k D,等电点为5.30,属于疏水性蛋白,可能定位于微体中。此外,艾纳香UFGT蛋白的二级结构中α螺旋占33.07%,延伸链占10.83%,无规则卷曲56.10%,与三级结构预测结果一致。本研究对于探究黔产艾纳香类黄酮物质的生物合成分子机制有一定的指导意义,并为将来类黄酮的生物合成提供帮助。

Blumea balsamifera is genuine medicinal materials of Guizhou Province, and its secondary metabolites have important pharmacologic actions, such as brass, blumeatin, etc.. Glucosyltransferase （UFGT） is a key enzyme in secondary metabolism pathways ofBlumea balsamifera. In this study, Blumea balsamifera produced on Luodian County of Guizhou Province was used as research materials. The complete cDNA sequence was cloned by RNA-seq and its bioinformatics was analyzed. The analysis results showed that the full-length of UFGT gene was 1 527 bp. UFGT gene could encode 508 amino acids. The protein molecular weight was 56.155 kD and its isoelectric point was 5.30. This protein was a hydrophobic protein and probably located in the microbody. Meanwhile, analysis of secondary structure indicated that UFGT of Blumea balsamifera contained alpha helixs （33.07%）, extended strands （10.83%）, random coils （56.10%）, and this result was consistent with the prediction of its three-dimensional structure. This study might have certain guiding significance for the biosynthesis mechanism of flavonoids in Blumea balsamifera, and would be helpful for the future biosynthesis.