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10 mM ethylene glycol bis (2-aminoethyl ether)-N, N, N’, N’,-tetraacetic acid (EGTA)

10mM乙二醇双(2-氨基乙基醚)-N,N,N'',N'', - 四乙酸(EGTA)

Company: Dojindo Molecular Technologies
Catalog#: 342-01314
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In vitro Phosphorylation Assay of Putative Blue-light Receptor Phototropins Using Microsomal and Plasma-membrane Fractions Prepared from Vallisneria Leaves
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2015-11-05
[Abstract]  An aquatic angiosperm Vallisneria (Alismatales: Hydrocharitaceae) has been used as an excellent experimental material over a century to study the light regulation of dynamic intracellular movements including chloroplast redistribution and cytoplasmic streaming (Senn, 1908; Seitz, 1987; Takagi, 1997). However, understanding of the molecular mechanisms lagged behind because of difficulty in applying modern techniques such as gene transformation to this plant. Especially, which kind of photoreceptors function in these intriguing responses has long been an unsolved topic. Recently, genes encoding plant-specific blue-light receptor phototropins were isolated in Vallisneria, for the first time from aquatic plants (Sakai et al., 2015). Phototropins were identified ... [摘要]  水生被子植物Vallisneria(Alismatales:Hydrocharitaceae)已经在一个世纪内用作研究动态细胞内运动的光调节的优良实验材料,包括叶绿体再分布和细胞质流(Senn,1908; Seitz,1987 ; Takagi,1997)。然而,对分子机制的理解落后,因为难以应用现代技术,如基因转化到这个植物。特别地,哪种类型的光感受器在这些有趣的反应中起作用长期以来一直是未解决的主题。最近,在水生植物中首次在Vallisneria中分离编码植物特异性蓝光受体光托品的基因(Sakai等人,2015)。光金蛋白首先被鉴定为拟南芥中下胚轴向往性的光感受器,并且现在已知调节许多反应,包括各种植物物种中的叶绿体光定位运动(Christie,2007)。光催化素主要位于质膜上,并且它们由蓝光诱导的自磷酸化是信号转导途径的关键步骤(Sakamoto和Briggs,2002; Kong等,2006; et al。,2013; Inoue et al。,2010)。在这里,我们描述了使用Vallisneria的原始微粒体和质膜富集级分的体外蛋白磷酸化测定的方案,其使得我们能够验证光催化素的存在并表征它们的自身磷酸化反应。在这些分析之后,Sakai等人(2015)提出 Vallisneria 光催化素介导高强度蓝光诱导的叶绿体回避反应。

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