Analysis of differentially expressed transcripts from planthopper-infested wild rice (Oryza minuta)
Analysis of differentially expressed transcripts from planthopper-infested wild rice (Oryza minuta)
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DOI：
10.1007/s00299-004-0905-9
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发表时间：
2005-04-01
影响因子：
6.2
通讯作者：
Shin, JS
中科院分区：
文献类型：
--
作者：
Cho, SK;Jung, KW;Shin, JS
利用抑制消减杂交技术结合镜像定向选择技术构建了一个以稻飞虱侵染的野生稻为材料的消减文库。为了筛选文库中差异表达的转录本，我们应用含有960个随机克隆的cDNA微阵列，使用从对照和稻飞虱侵染样品的mRNA制备的cDNA探针进行反向北方印迹分析。基于从一式三份进行的实验获得的信号强度和表达比率，我们选择了383个克隆。通过北方印迹分析验证随时间升高的表达水平和总体概况。虽然Southern杂交显示筛选出的基因在O. minuta和O. sativa的表达谱也有不同的模式。功能分类将所鉴定的转录物置于亚细胞定位、代谢和蛋白质命运的类别中。这些表达序列标签的存在意味着O.微生物对昆虫侵染的抗性不仅可以通过防御相关基因的表达升高来实现，而且还可以通过增强的代谢活性来实现。
A subtracted library was constructed from planthopper-infested wild rice (Oryza minuta) by suppression subtractive hybridization in combination with mirror orientation selection. To screen the differentially expressed transcripts in the library, we applied a cDNA microarray containing 960 random clones in a reverse Northern blot analysis using cDNA probes prepared from the mRNAs of control and planthopper-infested samples. On the basis of the signal intensities and expression ratios obtained from experiments performed in triplicate, we selected 383 clones. The elevated expression levels and overall profiles over time were verified by Northern blot analysis. Although Southern blot analysis showed similar copy numbers of the screened genes in O. minuta and O. sativa, it also revealed that the expression profiles had a different pattern . Functional categorization placed the identified transcripts in the categories of subcellular localization, metabolism, and protein fate. The presence of these expressed sequence tags implies that resistance of O. minuta to insect infestation can be achieved not only by an elevated expression of defense-related genes but also by enhanced metabolic activities.