陆地棉GhCYP1基因的分离及其表达分析

Isolation and Expression Pattern of GhCYP1 Gene in Upland Cotton

应用SSH-PCR技术,在筛选到抗旱相关基因的EST序列基础上,采用RACE技术,分离克隆到全长为801bp,含编码序列(CDS)为522 bp的棉花抗旱相关基因GhCYP1。氨基酸序列比对发现,该基因与拟南芥、水稻、人等的亲环蛋白基因高度同源,都具有由11个连续氨基酸所组成的单结构域亲环蛋白典型结构,且在保守区都具有PPIase活性所必需的3个氨基酸(R、F、H)以及与环孢素A(CsA)结合位点密切相关的色氨酸(W)。利用半定量RT-PCR对该基因的表达模式进行分析,发现其在棉花的叶、茎、根中均有表达,且在根中的表达量最高,暗示了该基因可能与植物根系吸水及矿物运输有关,从而在棉花适应干旱胁迫环境中发挥着关键作用。

In this study,based on the EST sequences of drought-resistant relative genes by employing SSH-PCR technique,a full length 801 bp cDNA of Cyclophilin-like gene was cloned from cotton（Gossypium hirsutum）by using RACE technique,which containing a 522 bp coding sequence（CDS）,and was designated as GhCYP1.Amino acid sequence alignment analysis showed that the GhCYP1 was high identitied with cyclophilin from Arabidopsis thaliana,Oryza sativa and Homo sapiens,all these cyclophilins containing a typical single-domain with a composition of 11 amino acids,three essential amino acids（R,F,H）for PPIase activity and a tryptophan（W）closely related to the site for cyclosporine A（CsA）binding.Semi-quantitative PCR analysis showed that the GhCYP1 was found with constitutive expression in the root,stem and leaf tissues,whereas the RNA transcriptional level in the root was highest,which suggested that this gene may be associated with uptaking water and transporting mineral,and played a pivotal role in the process of cotton adaptation to the drought environment.