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[目的]构建干旱和盐碱共胁迫的玉米cDNA文库,对细胞功能相关基因表达进行初步分析。[方法]提取处理玉米YQ7-96品系的雌雄分化期(12片叶龄)的混合组织(叶、茎和花芽)总RNA,运用SMART技术构建pDNR-LIB载体的cDNA文库,并随机挑选克隆进行EST序列的BlastX比对和MIPS归类分析。[结果]随机挑取3 027个cDNA克隆进行测序,94.45%的EST长度大于400 bp,共获得1 861个单基因簇,其中维持正常生理活动的基因占65.36%;参与细胞内运输、细胞内信号传导、细胞防御和细胞循环和DNA代谢过程的基因分别占9.26%、6.58%、2.63%和3.16%。[结论]在成功构建干旱和盐碱共胁迫的玉米cDNA文库的基础上,对产生的EST进行了测序和分析归类,筛选出了细胞功能相关EST,为后续研究奠定了基础。
[Objective] The research aimed to construct a cDNA library of maize co-stressed by drought and saline so as to conduct a preliminary analysis of the expression of cell function-related genes. [Method] Total RNA was extracted from the mixed tissues (leaves, stems and flower buds) of maize YQ7-96 during sexual differentiation (12 leaf ages). The cDNA library of pDNR-LIB vector was constructed by SMART technique and randomly selected clones BlastX alignment and MIPS classification analysis of EST sequences. [Results] A total of 3 027 cDNA clones were randomly selected and sequenced. 94.45% of the ESTs were over 400 bp in length. A total of 1 861 single gene clusters were obtained, of which 65.36% were normal physiological activities. They were involved in intracellular trafficking, intracellular The genes of signal transduction, cell defense and cell cycle and DNA metabolism accounted for 9.26%, 6.58%, 2.63% and 3.16% respectively. [Conclusion] Based on the successful construction of a cDNA library of drought and saline-alkali co-maize, the ESTs were sequenced and analyzed, and the ESTs of the cell function were screened out, which laid the foundation for subsequent research.