用SSH构建中滇苜蓿叶片文库及热胁迫相关基因初步分析

Yong-Gang Ding, W. Chang, Shi-Bao Zhang, Hong Hu
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摘要

高温可能是中滇incarvilla zhongdiannensis引种驯化到低海拔地区的主要限制因素之一。为了进一步揭示中滇稻对高温响应的潜在分子机制,本研究采用抑制减法杂交方法(SSH)进行研究。结果表明,正反相减cDNA文库质量良好,可用于进一步的SSH实验。通过SSH和测序随机获得60个高质量表达标签(est)。根据BLAST筛选和功能标注,将50条已知功能的est分为防御/胁迫、光合作用、代谢、能量、信号转导和转录、蛋白质命运、转运体、生物合成、细胞结构和细胞生长等9大类。有6个ESTs与假设的蛋白质显著匹配。4条无命中的ESTs可能编码了新的基因,在中滇家蚕应对高温胁迫中发挥了重要作用。
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Construction of Leave Library by SSH and Preliminary Analysis of Genes Responsible for Heat Sress in Incarvillea zhongdiannensis
High temperature might be one of the main limiting factors to be introduced and domesticated into lower altitude for Incarvillea zhongdiannensis. To further reveal the underlying molecular mechanism in I.zhongdiannensis responding to high temperature, the suppression subtractive hybridization approach (SSH) was used in this study. The results showed the quality of forward or reverse subtraction cDNA library is good enough for further SSH experiments. Total sixty expressed tags (ESTs) with high quality were randomly obtained by SSH and sequencing. According to BLAST screening and functional annotation, 50 ESTs with known function were classified into 9 broad categories such as defense/stress, photosynthesis, metabolism, energy, signal transduction and transcription, protein fate, transporter, biosynthesis, cell structure and cell growth. There were six ESTs significantly matched with hypothetical proteins. Four ESTs with no hits possibly encode novel genes and play important role in I.zhongdiannensis responding to high temperature stress.
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