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首页> 美国卫生研究院文献>Plant Physiology >DNA Sequence-Based Bar Codes for Tracking the Origins of Expressed Sequence Tags from a Maize cDNA Library Constructed Using Multiple mRNA Sources
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DNA Sequence-Based Bar Codes for Tracking the Origins of Expressed Sequence Tags from a Maize cDNA Library Constructed Using Multiple mRNA Sources

机译：基于DNA序列的条形码用于跟踪使用多个mRNA源构建的玉米cDNA文库中表达的序列标签的来源

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摘要

To enhance gene discovery, expressed sequence tag (EST) projects often make use of cDNA libraries produced using diverse mixtures of mRNAs. As such, expression data are lost because the origins of the resulting ESTs cannot be determined. Alternatively, multiple libraries can be prepared, each from a more restricted source of mRNAs. Although this approach allows the origins of ESTs to be determined, it requires the production of multiple libraries. A hybrid approach is reported here. A cDNA library was prepared using 21 different pools of maize (Zea mays) mRNAs. DNA sequence “bar codes” were added during first-strand cDNA synthesis to uniquely identify the mRNA source pool from which individual cDNAs were derived. Using a decoding algorithm that included error correction, it was possible to identify the source mRNA pool of more than 97% of the ESTs. The frequency at which a bar code is represented in an EST contig should be proportional to the abundance of the corresponding mRNA in the source pool. Consistent with this, all ESTs derived from several genes (zein and adh1) that are known to be exclusively expressed in kernels or preferentially expressed under anaerobic conditions, respectively, were exclusively tagged with bar codes associated with mRNA pools prepared from kernel and anaerobically treated seedlings, respectively. Hence, by allowing for the retention of expression data, the bar coding of cDNA libraries can enhance the value of EST projects.
机译：为了增强基因发现，表达的序列标签（EST）项目通常利用利用多种mRNA混合物产生的cDNA文库。这样，表达数据丢失，因为不能确定所得EST的来源。或者，可以制备多个文库，每个文库都来自更严格的mRNA来源。尽管此方法可以确定EST的来源，但它需要生成多个库。这里报道了一种混合方法。使用21个不同的玉米（Zea mays）mRNA集合制备cDNA文库。在第一链cDNA合成过程中添加了DNA序列“条形码”，以唯一鉴定衍生出单个cDNA的mRNA来源库。使用包含错误校正的解码算法，可以识别出超过97％的EST的源mRNA池。条形码在EST重叠群中出现的频率应与源库中相应mRNA的丰度成正比。与此相一致，所有分别由几个基因（玉米蛋白和adh1）衍生的所有EST（分别已知在核中专门表达或在厌氧条件下优先表达）分别用与从核和厌氧处理的幼苗制备的mRNA库相关的条形码专门标记。 ， 分别。因此，通过允许保留表达数据，cDNA文库的条形码可提高EST项目的价值。

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