陆地棉干旱胁迫响应基因GhGR的克隆及特征分析

【目的】克隆陆地棉干旱胁迫谷胱甘肽还原酶基因（GhGR），并对其序列进行生物信息学分析和表达分析。【方法】利用RACE和RT-PCR技术克隆陆地棉谷胱甘肽还原酶基因的全长序列，应用生物信息学软件对获得的基因序列及编码的蛋白序列进行分析；通过基因枪转化和实时荧光定量PCR表达对该基因表达部位和表达模式进行分析。【结果】从陆地棉（Gossypium hirsutum L.）中克隆了谷胱甘肽还原酶基因GhGR，cDNA全长1 035 bp，其中，ORF为792 bp，编码263个氨基酸。氨基酸序列比对和同源性分析显示该基因与杨树（XP_002299276.1）、蓖麻（XP_002518118.1）、葡萄（CAN74593.1）同源性最高，分别为90%、91%和91%。系统发育树结果显示，GhGR与葡萄中该蛋白的亲缘关系最近。基因枪转化和实时荧光定量PCR分析表明GhGR定位于洋葱的细胞膜和细胞核膜，并且其表达量受干旱胁迫诱导上调表达。【结论】从陆地棉克隆得到谷胱甘肽还原酶基因GhGR，初步认为该基因对干旱胁迫有一定响应。

Cloning and Characterization of Drought-stress Responsive Gene GhGR in Gossypium hirsutum L.

【Objective】The objective of this study is to clone and make bioinformatics analysis of the sequence of the glutathione reductase gene from G.hirsutum L.【Method】The full length of GhGR was cloned by RACE and RT-PCR techniques,the gene sequence obtained and the putative amino acid sequence were analyzed by bioinformatics software.Its expression patterns were analyzed by real-time PCR and instantaneous expression.【Result】The GhGR full length cDNA sequence contained 1 035 bp,and ORF was 792 bp,which encoded 263 amino acid residues.Bioinformatics analysis showed that GhGR had high similarity of 90%,91% and 91% to Populus trichocarpa（XP₀02299276.1）,Ricinus communis（XP₀02518118.1） and Vitis vinifera（CAN74593.1）,respectively.Phylogenetic analysis showed that GhGR was much closer to Vitis vinifera,the expression protein of GhGR was located in the cell membrane and cell nucleus membrane,and its expression could be induced by drought.【Conclusion】 The glutathione reductase gene GhGR was cloned for the first time from G.hirsutum L.The further studies of GhGR will have an important significance in drought-tolerance for cotton breeding.