摘要
以苋菜试管苗为材料,利用实验室构建的苋菜转录组数据库,筛选出WRKY13基因cDNA序列,对其开放阅读框(ORF)进行克隆验证和生物信息学分析。结果表明,WRKY13基因ORF长度为675 bp,编码224个氨基酸。通过生物信息学预测,该蛋白属于疏水性蛋白,有44个氨基酸位点存在磷酸化修饰;是WRKY蛋白超级家族(WRKY superfamily)的一个成员,存在1个WRKY结合位点;其二级结构由无规则卷曲结构、β折叠组成和α螺旋;蛋白系统进化树分析表明苋菜WRKY13蛋白独立形成一个分支,但与拟南芥、芜菁、烟草、葡萄聚类到一起,说明WRKY13蛋白在生物进化过程中还是比较保守的。本研究获得了苋菜WRKY13基因完整ORF,这为将来深入研究苋菜WRKY13基因功能提供了科学依据,也为研究其调控甜菜红素代谢过程的影响提供参考。
The in vitro plantlets of Amaranthus tricolor were used as the material. The c DNA sequence of WRKY13 gene was screened from transcriptome database of A. tricolor constructed in the laboratory, and the open reading frame(ORF) was used for cloning and bioinformatic analysis. The result showed that the WRKY13 ORF sequence was 675 bp in length, encoding 224 amino acids. Bioinformatics analysis showed that the protein was hydrophobic protein, and 44 amino acid sites were with phosphorylation modification. The protein was a member of WRKY superfamily with a WRKY binding site. The secondary structure consisted of random coil structure,β-sheet, and alpha helix. The analysis of protein phylogenetic tree indicated that WRKY13 protein of A. tricolor constituted a branch alone, but was associated with Arabidopsis thaliana, Brassica rapa, Nicotiana attenuata and Vitis vinifera, which indicated WRKY13 protein possesses relative conservatism during species evolution. This study has obtained the complete ORF of WRKY13 in A. tricolor, which would be helpful for further study of WRKY13 functions in A. tricolor, and might provide a reference for the study of the effects of A. tricolor WRKY13 genes on the metabolic processes of betacyanin.
引文
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