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【目的】分离土壤干旱胁迫条件下玉米幼苗期叶片诱导表达的基因。【方法】本研究利用抑制差减杂交(suppression subtractive hybridization,SSH)的方法,以耐旱自交系CN165为材料,构建了土壤干旱胁迫下玉米幼苗叶片的正向抑制差减cDNA文库。【结果】在这个文库中,随机挑取了672个阳性克隆,并进行PCR验证,对所有的单克隆进行了测序。得到了598个有效序列,经过EST聚类分析后,共得到了80个uniESTs。其中57个为contigs。23个为singlets。BLASTN的结果表明,所有的uniESTs都可以在玉米的核酸数据库中找到同源序列。BLASTX的结果表明:68个uniESTs和已知功能的蛋白有高度的相似性,8个uniESTs为未知功能蛋白和假定蛋白,4个uniESTs没有蛋白质的相似性。【结论】在这个cDNA文库中发现了大量抗旱相关的基因,这些基因涉及到植物代谢的多种途径。
【Objective】 The purpose of this study was to isolate the genes induced by leaves of maize seedlings under soil drought stress. 【Method】 In this study, suppression subtractive hybridization (SSH) method and drought tolerance inbred line CN165 were used to construct a subtractive cDNA library of maize seedlings under soil drought stress. [Results] In this library, 672 positive clones were randomly selected and subjected to PCR validation. All of the clones were sequenced. 598 valid sequences were obtained. After the EST clustering analysis, a total of 80 uniESTs were obtained. 57 of them are contigs. 23 for singlets. The BLASTN results show that all uniESTs can find homologous sequences in the corn nucleic acid database. The results of BLASTX showed that 68 uniESTs had a high degree of similarity with known proteins, 8 uniESTs were unknown and hypothetical, and 4 uniESTs did not have similar proteins. 【Conclusion】 A large number of drought-related genes were found in this cDNA library. These genes involved in various pathways of plant metabolism.