中间锦鸡儿WRKY30的克隆及生物信息学分析

Cloning and Bioinformatics Analysis of Caragana intermedia WRKY30

为了研究CiWRKY30转录因子在植物应对多种胁迫中发挥重要的作用,及对植物形态发育方面的影响。本研究以生长于内蒙古干旱高原地带且具有较高的抗寒抗旱特性的中间锦鸡儿为材料,从本实验室干旱转录数据库中获得胁迫响应转录因子CiWRKY30核苷酸序列,NCBI比对为全长序列,设计引物对该基因进行互补脱氧核糖核酸(complementary DNA,cDNA)克隆,测序结果显示CiWRKY30的开放阅读框(open reading frame,ORF)长1083 bp,编码361个氨基酸。然后通过生物信息学的比对分析方法,来对CiWRKY30和其它7种WRKY转录因子的分子量、等电点等一级结构,及其它二三级结构加以预测。结果推测这8种同源性较高蛋白很可能在细胞核中发挥作用,由此得到有效信息来对接下来的功能研究起到推动作用。另外,通过对CiWRKY30转录因子氨基酸序列进行生物信息学分析,预测CiWRKY30转录因子的结构,推断其可能的功能,以及对其他同源性较高植物WRKY转录因子结构功能的预测都有助于我们对CiWRKY30转录因子的研究加以深入了解。

In order to study the important role of CiWRKY30 transcription factor in plant response to various stresses and its effect on plant morphological development.Caragana intermedia,which grew in the arid plateau of Inner Mongolia and had high cold resistance and drought resistance,was used as material to obtain the nucleotide sequence of transcription factor CiWRKY30 from the drought transcription database of our laboratory,which is related with stress.NCBI alignment was used as the full-length sequence.A primer was designed to clone the gene and the result was the open reading frame(ORF)of CiWRKY30,which has 1 083 bp and encodes 361 amino acids.The molecular weight,isoelectric point and other secondary and tertiary structures of CiWRKY30 and seven other WRKY transcription factors,were predicted by bioinformatics comparative analysis.The results suggest that homologous elevated proteins in these 8 genes may play a role in the nucleus of the cells and thus provide effective information.It will help push the research down.In addition,bioinformatics analysis of the amino acid sequence of the transcription factor CiWRKY30,prediction of the structure of the transcription factor CiWRKY30,inference of its possible function,and prediction of the structure and function of other higher homologous plants WRKY transcription factor would be helpful to further understand the transcription factor CiWRKY30.