摘要
[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B lastX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] To construct corn cDNA library under the co-stress of drought and salinity,and analyze initially the expression of gene which related with the cell function.[Method] The total RNA of mixture tissue（leaf,stem and blossom bud） in the male and female differentiation period（12 leaves age） of corn YQ7-96 was extracted.By using SMART technology,cDNA library of pDNR-LIB vector was constructed.BlastX comparison and MIPS classification analysis of EST sequence were carried out by picking randomly the clone.[Result] 3 027 cDNA clones were picked out to sequence.94.45% EST length was bigger than 400 bp,and 1 861 single gene clusters were gained.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 constructing successfully corn cDNA library under co-stress of drought and salinity,the generated EST was sequenced,analyzed and classified.EST which related to the cell function was screened,which laid the foundation for the following research.
出处
《安徽农业科学》
CAS
北大核心
2011年第25期15413-15415,共3页
Journal of Anhui Agricultural Sciences
关键词
玉米
干旱
盐碱
共胁迫
EST
Corn
Drought
Saline and alkaline
Co-stress
EST
