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利用实时逆转录-聚合酶链反应对拟南芥类糖原合成酶激酶多基因家族进行表达谱分析。

Expression profiling of the whole Arabidopsis shaggy-like kinase multigene family by real-time reverse transcriptase-polymerase chain reaction.

作者信息

Charrier Bénédicte, Champion Anthony, Henry Yves, Kreis Martin

机构信息

Laboratoire de Biologie du Développement des Plantes, Bâtiment 630, Unité Mixte de Recherche-Centre National de la Recherche Scientifique 8618, Université Paris-Sud (XI), 91405 Orsay cedex, France.

出版信息

Plant Physiol. 2002 Oct;130(2):577-90. doi: 10.1104/pp.009175.

Abstract

The recent publication of the complete sequence of the Arabidopsis genome allowed us to identify and characterize the last two members of the SHAGGY-like kinase (AtSK) gene family. As a result, the study of the overall spatio-temporal organization of the whole AtSK family in Arabidopsis has become an achievable and necessary aim to understand the role of each SHAGGY-like kinase during plant development. An analysis of the transcript level of the 10 members of the family has been performed using the technique of real-time quantitative reverse transcriptase-polymerase chain reaction. Transcript levels in several organs, under different growth conditions, were analyzed. To calibrate the results obtained, a number of other genes, such as those coding for the two MAP3Kepsilons and the two MAP4Kalphas, as well as the stress response marker RD29A; the small subunit of the Rubisco photosynthetic enzyme Ats1A; the MEDEA chromatin remodeling factor; and the SCARECROW, ASYMMETRIC LEAVES 1, and SUPERMAN transcription factors all involved in key steps of plant development were used. The analysis of our data revealed that eight of the 10 genes of the AtSK family displayed a pseudo-constitutive expression pattern at the organ level. Conversely, AtSK13 responded to osmotic changes and saline treatment, whereas AtSK31 was flower specific and responded to osmotic changes and darkness.

摘要

拟南芥基因组完整序列的近期公布,使我们能够鉴定并描述类SHAGGY激酶(AtSK)基因家族的最后两个成员。因此,研究整个AtSK家族在拟南芥中的整体时空组织,已成为理解每个类SHAGGY激酶在植物发育过程中作用的一个可实现且必要的目标。利用实时定量逆转录-聚合酶链反应技术,对该家族10个成员的转录水平进行了分析。分析了在不同生长条件下几个器官中的转录水平。为校准所得结果,使用了许多其他基因,如编码两个MAP3Kε和两个MAP4Kα的基因,以及应激反应标记物RD29A;光合酶Rubisco的小亚基Ats1A;染色质重塑因子MEDEA;以及参与植物发育关键步骤的稻草人、不对称叶片1和超人转录因子。对我们数据的分析表明,AtSK家族10个基因中的8个在器官水平上呈现假组成型表达模式。相反,AtSK13对渗透变化和盐处理有反应,而AtSK31具有花特异性,对渗透变化和黑暗有反应。

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