中国农业科学2012，Vol.45Issue(2)：302-310,9.DOI:10.3864/j.issn.0578-1752.2012.02.012
番茄水通道蛋白基因SIAQP的克隆与序列分析
Cloning and Sequence Analysis of the Aquaporins Gene SIAQP in Tomato
摘要
Abstract
[Objective] To provide basal data for mechanism of tomato's drought resistance and cultivar development, the sequence characteristics of the aquaporin gene S1AQP in tomato were analyzed, the coded protein was intracellar located and the expression profiling of the Mirco Tom tomato was studied after treatment under drought stress. [Method] The rapid-amplification of cDNA ends (RACE) was used to amplify the full-length S1AQP gene, and the bioinformatics software was used to analyze the structures and function of the coded protein. In order to follow the intracellular localisation of the protein, the GFP sequence was fused downstream to the S1AQP coding region and the fusion gene S1AQP::GFP transferred into onion epidermal cells by biolistic method. The Mirco Tom tomato was used as material, the real time-PCR was adopted to study the expression profile of gene S1AQP. [Result] The full-length cDNA of S1AQP gene (GenBank Accessin No. HQ433337) consists of 1 107 bp and contains a 852 bp open reading frame (ORF) encoding 283 amino acid proteins. Bioinformatics analysis demonstrated that S1AQP gene exhibited a typical structure with six membrane-spanning domains and an internal symmetry showing two highly conserved Asn-Pro-Aja (NPA) motifs, and possessing the MIP family signal consensus sequence. The S1AQP amino acids showed high identity with other 10 plantspecies PIP subfamily by NCBI homology comparison analysis. Phylogenetic analysis among 11 species indicated that S1AQP was clustered with the StAQP from Solarium tuberosum. The result of transient expression showed that S1AQP gene was located in the membrane. Southern blot analysis indicated that SIAQP was a single-copy gene. Real time-PCR analysis showed that the expression of SIAQP gene was down-regulated by drought stress. [Conclusion] The expression profiling of this gene indicates that the SIAQP gene may regulared by drought stress. This paper provides important information for the future study on the gene-expression regulation during drought stress.关键词
番茄/水通道蛋白/序列分析/细胞定位/Real time-PCRKey words
tomato/ aquaporins/ sequence analysis/ intracellular localization/ Real time-PCR引用本文复制引用
李仁,吴新新,李蔚,杨荣超,赵永钦,温常龙,赵冰,郭仰东..番茄水通道蛋白基因SIAQP的克隆与序列分析[J].中国农业科学,2012,45(2):302-310,9.基金项目
国家重点基础研究发展计划项目(2009CB119000)、国家自然科学基金项目(31171989)、中央高校基本科研业务费专项(2009-2-06) （2009CB119000）
