In Gel Kinase Assay
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Author:
Date:
2017-03-05
[Abstract] Proper spatiotemporal regulation of protein phosphorylation in cells and tissues is required for normal development and homeostasis. We present the protocol ‘In Gel Kinase Assay’, which is useful for protein kinase activity measurements from crude protein extracts. We have successfully used ‘In Gel Kinase Assay’ protocol to show that the Arabidopsis thaliana sextuple mutant in the PYRABACTIN RESISTANCE1/PYR1-LIKE/REGULATORY COMPONENTS OF ABA RECEPTORS (PYR/PYL/RCAR-ABA receptors; line pyr/pyl112458) is impaired in ABA-mediated activation of SnRK2.2, SnRK2.3 and OST1/SnRK2.6, ...
[摘要] 正常发育和体内平衡需要细胞和组织中蛋白质磷酸化的适时时空调节。我们提出方案“凝胶激酶测定”,其可用于粗蛋白质提取物的蛋白激酶活性测量。我们已经成功地使用“凝胶激酶测定”方案来证明在ABA受体(PYR / PYL / RCAR-ABA受体)的PYRABACTIN RESISTANCE1 / PYR1-样/调节组分中的拟南芥线条pyr / pyl112458 )在ABA介导的SnRK2.2,SnRK2.3和OST1 / SnRK2.6的活化中受损,多达三重突变体snrk2.2 / 2.3 / 2.6 (Gonzalez-Guzman等人,2012)。
背景 植物激素脱落酸(ABA)是涉及植物生长发育以及植物对非生物和生物胁迫的反应的关键信号。 ABA感知和信号通路由ABA受体(PYR / PYL / RCAR-ABA受体)的PYRABACTIN RESISTANCE1 / PYR1-调节组分,PP2C磷酸酶和SnRK2s激酶组成(在Antoni等人, ,,2011)。模块受体-ABA-磷酸酶通过调节ABA激活的SnRK2而以配体依赖的方式控制磷酸化信号级联。反过来,SnRK2s激酶使细胞核和细胞质中的无数效应物从转录因子(例如,ABFs)到离子通道(例如)磷酸化, ...
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Primer Extension Analysis Using AMV Reverse Transcriptase
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Author:
Date:
2012-07-20
[Abstract] Primer extension analysis is a useful method to determine the transcription start point or a processing site on an RNA molecule. It can also allow a quantitative measurement of an RNA species.
[摘要] 该实验方案的中文版正在准备中...
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Synthesis of 5’ end-labeled RNA
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Author:
Date:
2012-07-20
[Abstract] 5’ end-labeled RNA molecules are useful substrates to analyse the endo- and exonucleolytic activities of various ribonucleases. Here two protocols are given to synthesize P32 labeled RNAs with a 5’ PPP or 5’ P moiety. 5’ exoribonucleases generally do not work on 5’ PPP RNA and require a 5’ P substrate. The activity of certain endoribonucleases like Escherichia coli (E. coli) RNase E or Bacillus subtilis (B. subtilis) RNase Y can be stimulated by a 5’ P moiety.
[摘要] 该实验方案的中文版正在准备中...
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