| Self-organization Assay for Min Proteins of Escherichia coli in Micro-droplets Covered with Lipids
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Author:
Date:
2020-03-20
[Abstract] The Min system determines the cell division plane of bacteria. As a cue of spatiotemporal regulation, the Min system uses wave propagation of MinD protein (Min wave). Therefore, the reconstitution of the Min wave in cell-sized closed space will lead to the creation of artificial cells capable of cell division. The Min waves emerge via coupling between the reactions among MinD, MinE, and ATP and the differences in diffusion rate on the cell membrane and in the cytoplasm. Because Min waves appear only under the balanced condition of the reaction-diffusion coupling, special attentions are needed towards several technical points for the reconstitution of Min waves in artificial cells. This protocol describes a technical method for stably generating Min waves in artificial cells.
[摘要] [摘要 ] Min系统确定细菌的细胞分裂平面。作为时空调节的提示，Min系统使用MinD 蛋白的波传播（Min wave）。因此，Min波在细胞大小的封闭空间中的重构将导致能够分裂细胞的人造细胞的产生。闵波出现经由耦合之间反应小号中MinD的，的MinE ，和ATP 和所述differenc ES 在细胞膜上的扩散速度和在细胞质中。因为最小波仅在反应扩散耦合的平衡条件下出现， 特别关注，需要对几个技术要点为闽波在人造细胞重建。该协议描述了一种在人造细胞中稳定产生Min波的技术方法。
[背景 ] 敏系统，它决定了细胞分ER 对称细胞分裂，是在细菌细胞内的组织系统的最显着的例子之一（Rothfield 等人，2005;和罗利特马戈林，2013年）。敏系统使用图案形成在细胞内的时间依赖性蛋白梯度的公知的作为敏波（宽松等人，2008; Halatek和Frey，2012;邦尼等人，2013; Zieske 。等人，2016 ; Kohyama 。等人， 2019 ）。Min波是由两种蛋白MinD 和MinE 的反应扩散耦合产生的。通过与ATP结合，MinD 形成二聚体并附着在膜上。的MinE 被招募到的ATP MinD的和诱导ATP酶的活性MinD的。通过MinE ，ATP- MinD 变为ADP- MinD ，并从膜上脱离。ADP- MinD的被转换回ATP- ...
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| Incubation of Cyanobacteria under Dark, Anaerobic Conditions and Quantification of the Excreted Organic Acids by HPLC
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Author:
Date:
2017-05-05
[Abstract] Succinate and lactate are commodity chemicals used for producing bioplastics. Recently, it was found that such organic acids are excreted from cells of the unicellular cyanobacterium Synechocystis sp. PCC 6803 under dark, anaerobic conditions. To conduct the dark, anaerobic incubation, cells were concentrated within a vial that was then sealed with a butyl rubber cap, following which N2 gas was introduced into the vial. The organic acids produced were quantified by high-performance liquid chromatography via post-labeling with bromothymol blue as a pH indicator. After separation by ion-exclusion chromatography, the organic acids were identified by comparing their retention time with that of standard solutions. These procedures allow researchers to quantify the organic ...
[摘要] 琥珀酸盐和乳酸盐是用于生产生物塑料的商品化学品。最近发现这样的有机酸从单细胞蓝细菌集胞藻的细胞中排出。 PCC 6803在黑暗，无氧条件下。为了进行黑暗的厌氧培养，将细胞浓缩在小瓶内，然后用丁基橡胶帽密封，随后将N 2气体引入小瓶中。通过用溴百里酚蓝作为pH指示剂的后标记，通过高效液相色谱定量产生的有机酸。通过离子排阻色谱分离后，通过将其保留时间与标准溶液的保留时间进行比较来鉴定有机酸。这些程序允许研究人员量化微生物产生的有机酸，有助于蓝藻生物学和生物技术的知识。
背景 蓝细菌是一组进行氧合光合作用的细菌。集胞藻 sp。 PCC 6803（以下称为集胞藻6803）是非固氮，单细胞蓝细菌，通常用于基础和应用研究。集胞藻6803细胞能够在黑暗和厌氧条件下分泌有机酸，如琥珀酸盐和乳酸盐（Osanai等，2015）。培养条件的遗传操作和修饰，例如添加钾或NaHCO 3，成功地提高了从集胞藻6803细胞排出的有机酸的水平（Osanai >等人，2015; Hasunuma等人，2016; Iijima等人，2016; Ueda等人。 ...
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