Expression, Purification and Enzymatic Assay of Plant Histone Deacetylases
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Author:
Date:
2016-04-05
[Abstract] Histone deacetylases (HDACs) catalyzing the removal of acetyl groups from lysine residues of histone and non-histone proteins play vital roles in regulation of gene transcription. In plants, HDACs can be grouped into three families, including RPD3-type, SIR2-type and plant specific HD2-type HDACs. Here we describe a method to determine plant HDAC enzymatic activity. This protocol includes expression, purification and enzymatic activity assay of recombinant plant HDACs expressed in Escherichia coli (E. coli) and Arabidopsis thaliana (A. thaliana).
[摘要] 组蛋白脱乙酰酶(HDAC)催化从组蛋白和非组蛋白蛋白的赖氨酸残基去除乙酰基在调节基因转录中起重要作用。 在植物中,HDAC可以分为三个家族,包括RPD3型,SIR2型和植物特异性HD2型HDAC。 在这里我们描述了一种确定植物HDAC酶活性的方法。 该方案包括在大肠杆菌(大肠杆菌)和拟南芥()中表达的重组植物HDAC的表达,纯化和酶活性测定 >拟南芥)。
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Isolation of Chloroplast Inner and Outer Envelope Membranes
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Author:
Date:
2015-02-20
[Abstract] The chloroplast is an important organelle found in plant cells that conduct photosynthesis. It is enclosed by a pair of closely spaced membranes, the double-membrane envelope, consisting of the inner membrane bounding the matrix or stroma and the outer membrane in contact with the cytoplasm. Like many bio-membranes, the chloroplast envelope plays an important role in mediating the complex interactions between the chloroplast and the cytoplasm. The envelope is also the site of various biosynthetic reactions, including the formation of the galactolipids, which are the major components of both ...
[摘要] 叶绿体是在进行光合作用的植物细胞中发现的重要细胞器。它被一对紧密间隔的膜包围,双膜包膜由包围基质或基质的内膜和与细胞质接触的外膜组成。像许多生物膜一样,叶绿体包膜在介导叶绿体和细胞质之间的复杂相互作用中起重要作用。包膜也是各种生物合成反应的位点,包括半乳糖脂的形成,半乳糖脂是包膜和类囊体膜的主要成分。内封套膜和外封套膜在结构和功能上都有差异。例如,外膜表现出比内膜剂量更低的膜内颗粒密度,表明外膜的蛋白质含量低。此外,外膜对于低分子量化合物是非特异性可渗透的,而内部膜对于这些化合物是不可渗透的,并且包含用于运输代谢物的几种易位体系统。 为了制备包膜,必须首先分离完整的叶绿体。然后通过以下方式分离内外膜:1)蛋白酶处理方法和2)基于外壳更轻且内膜更重的事实的离心方法。两种方法都需要首先分离完整的叶绿体。然而,离心分离可以得到纯的内外封套制剂,因此适合于随后的分析。此外,离心法可以避免在蛋白酶处理期间内皮多肽的破坏,因为一些蛋白酶可以进入内膜。此外,离心方法容易操作并且获得包含较少外部和内部信封膜的粘合区域的完全包封。在这里,我们描述了一种可靠的方法,用于从烟草中分离叶绿体的内和外包膜,这是相对不容易用于包膜分离的植物。
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Preparation of Primary Neurons from Rat Median Preoptic Nucleus (MnPO)
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Author:
Date:
2013-12-05
[Abstract] Studying cell physiology is really important to understand their function and to determine action mechanisms taking place in these cells. Using brain slices can be sometime difficult to directly access to cells like neurons. Even if it more steps are needed to get cultured cells, this type of preparation allow a better access to neurons and provide a good way to study their basal properties. Here we describe a protocol of a short term primary neurons culture from MnPO, allowing to kept neurons in good condition to perform electrophysiological recordings.
[摘要] 研究细胞生理学对于了解其功能和确定在这些细胞中发生的作用机制是非常重要的。 使用脑切片可能有时难以直接访问像神经元这样的细胞。 即使需要更多的步骤来培养细胞,这种类型的制剂可以更好地接近神经元,并提供研究其基础性质的好方法。 在这里,我们描述了一种来自MnPO的短期原代神经元培养的方案,允许神经元保持良好的状态进行电生理记录。
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