Analysis of gene expression by ESTs from suppression subtractive hybridization library in Chenopodium album L. under salt stress
Analysis of gene expression by ESTs from suppression subtractive hybridization library in Chenopodium album L. under salt stress
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DOI：
10.1007/s11033-011-0678-5
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发表时间：
2011-11-01
影响因子：
2.8
通讯作者：
Zhang, Fuchun
中科院分区：
文献类型：
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作者：
Gu, Lili;Xu, Dongsheng;Zhang, Fuchun
为鉴定盐胁迫下藜基因的表达，筛选与盐胁迫相关的EST，利用消减抑制杂交（SSH）技术构建了盐胁迫下藜基因文库。本研究构建了盐胁迫条件下的白蛋白表达载体。随机EST测序共获得825条高质量的EST序列（GenBank ID为GE 746311-GE 747007），平均长度为301 bp，聚类为88个contig和550个singletons。根据功能注释将其分为12类。635条EST（76.97%）与非冗余数据库中的基因序列相似，190条EST（23.03%）与非冗余数据库中的基因序列相似性较低或无相似性。从SSH文库的347个未知或新的EST中随机选取56个EST，分析其在不同NaCl浓度和不同时间点的转录谱。结果表明，高比例的测试ESTs被盐胁迫激活。在56个对NaCl有反应的EST中，有4个在阿坝和PEG胁迫下表达量也增加。通过北方杂交验证了上述4条EST序列的正确性，与RT-PCR结果一致。结果表明，这些EST对应的基因可能参与胁迫反应或调控。这些EST的完整序列和详细功能有待进一步研究。
To identify genes expression in Chenopodium album exposed to NaCl stress and screen ESTs related to salt stress, a subtractive suppression hybridization (SSH) library of C. album under salt stress was constructed in the present study. Random EST sequencing produced 825 high-quality ESTs with GenBank ID GE746311-GE747007, which had 301 bp of average size and were clustered into 88 contigs and 550 singletons. They were classified into 12 categories according to their function annotations. 635 ESTs (76.97%) showed similarities to gene sequences in the non-redundancy database, while 190 ESTs (23.03%) showed low or no similarities. The transcriptional profiles of 56 ESTs randomly selected from 347 unknown or novel ESTs of SSH library under varying NaCl concentration and at different time points were analyzed. The results indicated that a high proportion of tested ESTs were activated by salt stress. Four in 56 ESTs responded to NaCl were also enhanced in expression level when exposed to ABA and PEG stresses. The above four ESTs were validated by northern blotting which was consistent with the results of RT-PCR. The results suggested that genes corresponded to these ESTs might be involved in stress response or regulation. The complete sequences and detailed function of these ESTs need to be further studied.