高温胁迫下不同玉米花粉组蛋白Zm-H3基因的克隆及表达分析

Cloning and Expression Analysis of Histone Zm-H3 Gene in Different Maize Pollen under High Temperature Stress

为了解组蛋白H 3基因(Zm-H3)在不同耐高温玉米(Zea mays L.)材料中的差异表达情况,以中地88(耐高温)和先玉335(不耐高温)2个玉米品种为材料,在高温条件下取其花粉提取总RNA,再通过mRNA纯化、反转录、文库构建及高通量测序等步骤,克隆了一个玉米组蛋白Zm-H3。生物信息学分析表明,该基因位于玉米第6染色体上,全长cDNA为904 bp,编码长度为136个氨基酸的蛋白质,进一步对该蛋白的理化性质、功能位点及结构作了分析。转录组高通量测序结果显示,Zm-H3基因在这2个玉米材料中的表达呈现显著差异,在中地88材料中出现了高表达,为先玉335材料的2.08倍。该实验结果可以为玉米组蛋白H 3基因的应用研究奠定分子基础,也可为其它基因的克隆及表达分析提供参考。

To understand differential expression of histone gene Zm-H3 in different high temperature-tolerant maizes,the two maize varieties Zhongdi 88(high temperature-tolerant variety)and Xianyu 335(high temperature-sensitive variety)were used as plant materials to cloned a histone Zm-H3 gene via series of steps including total RNA extract,mRNA purification,reverse transcription,library construction and high-throughput sequencing.Bioinformatics analysis showed that the Zm-H3 gene was located on chromosome 6 of maize with a full-length cDNA of 904 bp and encoded a protein with 136 amino acids.The physical and chemical properties,functional sites and structure of the protein were further analyzed.High-throughput transcriptome sequencing results showed that the expression of Zm-H3 gene was significantly different in the two maizes,with high expression in Zhongdi 88,which was 2.08 folds higher than that in Xianyu 335.The experimental results could lay a molecular foundation for the application of maize histone Zm-H3 gene,and also provided reference for the cloning and expression analysis of other genes.