日本蛇根草糖基转移酶基因Oj GT1的克隆与生物信息学分析

Cloning and Bioinformatics Analysis of OjGT1 Gene in Ophiorrhiza japonica

糖基转移酶(GTs)是花色苷合成过程中最重要的修饰酶之一,其在花色苷合成过程中发挥重要作用。本研究以日本蛇根草为材料,依据其转录组测序结果,设计引物通过RT-PCR方法成功克隆得到OjGT1基因完整的cDNA序列,随后对OjGT1的功能结构域、生理和化学参数、亲/疏水性、信号肽,二级结构等进行生物信息学分析。分析结果显示,OjGT1基因完整的CDS长度为1 413 bp,翻译形成蛋白的分子量为52.087 kD,等电点为5.12,编码形成亲水性蛋白质,不含信号肽。同时二级结构与三级结构分析显示,OjGT1蛋白结构主要以不规则卷曲为主,其次是α螺旋,延伸链所占比重最小。OjGT1基因的成功克隆与生物信息学分析,将为后续研究茜草目其它植物糖基转移酶提供参考,同时也为日本蛇根草花色苷的生物合成研究提供科学依据。

Glycosyltransferase(GTs) is a key modification enzyme in the anthocyanin biosynthetic pathway, and it plays an important role during anthocyanin biosynthesis. In this study, we used Ophiorrhiza japonica as materials, and designed the primers according to the results of its transcriptome sequencing. The integrated cDNA of OjGT1 was successfully cloned by RT-PCR method, and then it was applied to the bioinformatics analysis,including functional domains, physical and chemical parameters, hydrophobicity or hydrophilicity, signal peptide,secondary structure and so on. The results showed that the CDS of OjGT1 was 1 413 bp, the molecular weight of the translated protein was 52.087 kD, and its isoelectric points was 5.12. OjGT1 of Ophiorrhiza japonica was hydrophilic protein, which had no signal peptide. Analysis of secondary and tertiary structure indicated that OjGT1 protein was mainly composed of random coils, followed by the α-helix, and the extended strands occupied the smallest proportion. Successful cloning and bioinformatics analysis of OjGT1 could provide theoretical reference for the study on glycosyltransferase from other plants of Rubiales, and lay a foundation for further research on thebiosynthesis of anthocyanins in Ophiorrhiza japonica as well.