摘要
[Objective] The research aimed to construct maize cDNA library under the co-stress of drought,salt and alkali,and initially analyze the expression of gene which related to the cellular function.[Method] The total RNA of mixed tissue(leaf,stem and blossom bud) of maize YQ7-96 in the male and female differentiation period(12-leaf age) was extracted.By using SMART technology,cDNA library of pDNR-LIB vector was constructed.BlastX contrast and MIPS classification analysis of EST sequence were carried out by randomly picking the clone.[Result] 3 027 cDNA clones were picked out to sequence.The length of 94.45% EST was bigger than 400 bp,and 1 861 single gene clusters were obtained.The gene which maintained the normal physiological activity occupied 65.36%.The genes which involved in the intracellular transportation,signal transduction,cell defense and cycle,DNA metabolic process were respectively 9.26%,6.58%,2.63% and 3.16%.[Conclusion] Based on the successful construction of maize cDNA library under the co-stress of drought,salt and alkali,EST was sequenced,analyzed and classified.EST which related to the cellular function was screened.It laid the foundation for the following research.
[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。