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一个用于2500个水稻转录因子高通量表达谱分析的定量逆转录聚合酶链反应平台。

A quantitative RT-PCR platform for high-throughput expression profiling of 2500 rice transcription factors.

作者信息

Caldana Camila, Scheible Wolf-Rüdiger, Mueller-Roeber Bernd, Ruzicic Slobodan

机构信息

Max-Planck Institute of Molecular Plant Physiology, Am Mühlenberg 1, 14476 Potsdam-Golm, Germany.

出版信息

Plant Methods. 2007 Jun 8;3:7. doi: 10.1186/1746-4811-3-7.

Abstract

BACKGROUND

Quantitative reverse transcription - polymerase chain reaction (qRT-PCR) has been demonstrated to be particularly suitable for the analysis of weakly expressed genes, such as those encoding transcription factors. Rice (Oryza sativa L.) is an important crop and the most advanced model for monocotyledonous species; its nuclear genome has been sequenced and molecular tools are being developed for functional analyses. However, high-throughput methods for rice research are still limited and a large-scale qRT-PCR platform for gene expression analyses has not been reported.

RESULTS

We established a qRT-PCR platform enabling the multi-parallel determination of the expression levels of more than 2500 rice transcription factor genes. Additionally, using different rice cultivars, tissues and physiological conditions, we evaluated the expression stability of seven reference genes. We demonstrate this resource allows specific and reliable detection of the expression of transcription factor genes in rice.

CONCLUSION

Multi-parallel qRT-PCR allows the versatile and sensitive transcriptome profiling of large numbers of rice transcription factor genes. The new platform complements existing microarray-based expression profiling techniques, by allowing the analysis of lowly expressed transcription factor genes to determine their involvement in developmental or physiological processes. We expect that this resource will be of broad utility to the scientific community in the further development of rice as an important model for plant science.

摘要

背景

定量逆转录 - 聚合酶链反应(qRT-PCR)已被证明特别适用于分析弱表达基因,例如那些编码转录因子的基因。水稻(Oryza sativa L.)是一种重要的作物,也是单子叶植物中最先进的模式植物;其核基因组已被测序,并且正在开发用于功能分析的分子工具。然而,用于水稻研究的高通量方法仍然有限,尚未报道用于基因表达分析的大规模qRT-PCR平台。

结果

我们建立了一个qRT-PCR平台,能够多平行测定2500多个水稻转录因子基因的表达水平。此外,我们使用不同的水稻品种、组织和生理条件,评估了七个内参基因的表达稳定性。我们证明了该资源能够特异性且可靠地检测水稻中转录因子基因的表达。

结论

多平行qRT-PCR能够对大量水稻转录因子基因进行通用且灵敏的转录组分析。这个新平台通过允许分析低表达的转录因子基因以确定它们在发育或生理过程中的作用,补充了现有的基于微阵列的表达分析技术。我们期望该资源在将水稻进一步发展成为植物科学的重要模式植物的过程中,对科学界具有广泛的用途。

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