A quantitative RT-PCR platform for high-throughput expression profiling of 2500 rice transcription factors.
A quantitative RT-PCR platform for high-throughput expression profiling of 2500 rice transcription factors.
复制标题
DOI：
10.1186/1746-4811-3-7
复制
发表时间：
2007-06-08
期刊：
影响因子：
5.1
通讯作者：
Ruzicic S
中科院分区：
文献类型：
--
作者：
Caldana C;Scheible WR;Mueller-Roeber B;Ruzicic S
定量逆转录-聚合酶链反应（qRT-PCR）已被证明特别适用于弱表达基因的分析，如编码转录因子的基因。水稻（Oryza sativa L.）是重要的单子叶植物，也是最先进的单子叶植物模式；其核基因组已被测序，用于功能分析的分子工具正在开发中。然而，水稻研究的高通量方法仍然有限，用于基因表达分析的大规模qRT-PCR平台尚未报道。我们建立了一个qRT-PCR平台，可以多平行测定2500多个水稻转录因子基因的表达水平。此外，利用不同的水稻品种、组织和生理条件，对7个内参基因的表达稳定性进行了评价。我们证明这种资源允许特异性和可靠的检测转录因子基因在水稻中的表达。多重平行qRT-PCR允许对大量水稻转录因子基因进行多功能和敏感的转录组分析。新平台补充了现有的基于微阵列的表达谱技术，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参与。我们期望这一资源在进一步开发水稻作为植物科学的重要模型方面具有广泛的应用价值。
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. 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. 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.
登录
查看更多内容
影响因子：
6.9
作者：
Suzuki, Y;Makino, A;Mae, T
通讯作者：
Mae, T
影响因子：
4.1
作者：
Rajeevan, MS;Vernon, SD;Unger, ER
通讯作者：
Unger, ER
影响因子：
4.4
作者：
Wang, Yulei;Barbacioru, Catalin;Hyland, Fiona;Xiao, Wenming;Hunkapiller, Kathryn L;Blake, Julie;Chan, Frances;Gonzalez, Carolyn;Zhang, Lu;Samaha, Raymond R
通讯作者：
Samaha, Raymond R
影响因子：
3
作者：
Riaño-Pachón DM;Ruzicic S;Dreyer I;Mueller-Roeber B
通讯作者：
Mueller-Roeber B
DOI：
10.1073/pnas.0602039103
发表时间：
2006-05-02
影响因子：
11.1
作者：
Wang, BB;Brendel, V
通讯作者：
Brendel, V