日本蛇根草花青素还原酶基因的克隆及其生物信息学分析

Cloning and Bioinformatics Analysis of ANR Gene in Ophiorrhiza japonica

花青素还原酶(anthocyanidin reductase, ANR)是原花青素合成途径中的关键酶,同时花青素还原酶对植物花色苷的积累也发挥着重要作用。本研究以日本蛇根草为材料,以其转录组测序结果为基础,设计引物通过RT-PCR方法成功克隆得到日本蛇根草ANR基因完整的cDNA序列,利用生物信息学方法和工具对日本蛇根草ANR基因的功能结构域、生理和化学参数、亲/疏水性、信号肽、二级结构等进行预测和分析。结果表明,该基因cDNA全长为1 020 bp,编码339个氨基酸,预测其蛋白分子量为36.37 kD,等电点为5.92,为亲水蛋白,不含信号肽序列。综上,本研究成功克隆获得日本蛇根草ANR基因并完成其生物信息学分析,研究结果将为解析该基因的功能奠定基础。

Anthocyanidin reductase(ANR) is the key enzyme in the procyanidins biosynthetic pathway, and it also plays an important role in the accumulation of anthocyanin in plants. In this study, we utilized Ophiorrhiza japonica as materials, and designed the primers according to the results of the transcriptome sequencing. The integrated cDNA sequence of ANR gene was cloned successfully by RT-PCR method in Ophiorrhiza japonica. Bioinformatics methods and tools were applied to the functional domains, physical and chemical parameters, hydrophobicity or hydrophilicity, signal peptide, secondary structure and other factors in ANR gene of Ophiorrhiza japonica for prediction and analysis. The results showed that the full length of the gene cDNA was 1 020 bp, which encoded 339 amino acids. The protein molecular weight was predicted to be 36.37 kD and the isoelectric points were 5.92, ANR gene of Ophiorrhiza japonica was a hydrophilic protein with no signal peptide sequence. In summary, we have successfully cloned the ANR gene from Ophiorrhiza japonica and finished its bioinformatics analysis, these results will lay a foundation for the analysis of the function of this gene.