Quantitation of Cytochromes b559, b6, and f, and the Core Component of Photosystem I P700 in Cyanobacterial Cells
|
Author:
Date:
2016-11-05
[Abstract] Cytochrome (Cyt) b559, an important and essential core component of photosystem II in the photosynthetic electron transport system, is a heme-bridged heterodimer protein composed of an alpha subunit (PsbE) and a beta subunit (PsbE), and its reduced form has an absorption maximum in the α-band at 559 nm. The amounts of Cyt b559 can be determined by spectrophotometrical measurement of reduced minus oxidized difference spectra that are normalized with absorbance of isosbestic point at 580 nm. The authors use differential extinction ...
[摘要] dA 6m DNA免疫沉淀随后深度测序(DIP-Seq)是鉴定和研究N 6 - 甲基脱氧腺苷(dA 6)的全基因组分布的关键工具, 6m )。这种新的DNA修饰的精确功能仍然需要完全阐明,但是已知转录起始位点不存在并且从外显子中排除,表明在转录调节中的作用(Koziol等人,2015 )。重要的是,其存在表明DNA可能比先前认为的更多样化,因为进一步的DNA修饰可能存在于真核DNA中(Koziol等人,2015)。该方案描述了进行dA 6m DNA免疫沉淀(DIP)的方法,其用于表征高等真核生物中的第一dA 6m甲基化酶分析(Koziol等人。,2015)。在该协议中,我们描述了如何基因组DNA被分离,片段化,然后用识别基因组DNA中的dA 6m 的抗体拉下含有dA 6m 的DNA。在随后的洗涤之后,消除不含dA 6m的DNA片段,并且从抗体洗脱含有dA 6m的片段,以便进一步处理用于随后的分析。 [背景] 此协议是为了识别基因组中包含dA 6m 的区域而开发的。它可以用于检测不同基因组中的dA 6m 。作为指导,本方案从用于检测RNA中腺苷甲基化的现有方法建立(Dominissini等人,2013)。我们开发这个协议,并适应它的dA 6 m 在DNA中的检测,而不是检测腺苷甲基化RNA。这是必需的,因为当时没有方案可用于允许在真核DNA中dA ...
|
|
Determination of Hydroquinone Dioxygenase Activity
|
Author:
Date:
2015-09-05
[Abstract] We recently demonstrated the presence of a quinone detoxification pathway present in Firmicutes. It is based on two enzyme activities, namely a hydroquinone dioxygenase, YaiA, described here, and a hydroquinone reductase, YaiB, described in a separate protocol. In Lactococcus lactis (L. lactis), these enzymes are encoded by the yahCD-yaiAB operon. The operon is induced by copper to prevent the synergistic toxicity of quinones and copper. The hydroquinone dioxygenase, YaiA, converts hydroquinones to 4-hydroxymuconic semialdehyde, using molecular oxygen as oxidant ...
[摘要] 我们最近展示了存在于Firmicutes中的醌解毒途径。 它基于两种酶活性,即本文所述的氢醌双加氧酶,YaiA和在单独方案中描述的氢醌还原酶,YaiB。 在乳酸乳球菌(乳酸乳杆菌)中,这些酶由yahCD-yaiAB 操纵子编码。 操纵子由铜诱导以防止醌和铜的协同毒性。 根据以下反应,氢醌双加氧酶YaiA使用分子氧作为氧化剂将氢醌转化为4-羟基粘康酸半醛:氢醌+ O 2→4-羟基粘康酸半醛+ H + 我们在这里描述了用于测量氢醌双加氧酶活性的两种方法,基于用氧电极测量的氧消耗和4-羟基粘康酸半醛的分光光度检测。 两种测定用粗细胞提取物进行。
|
|