| Purification and Detection of a PDGA Depolymerase from Pusillimonas noertemannii
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Author:
Date:
2014-11-05
[Abstract] The purification of a target protein from a complex mixture of proteins is a challenging undertaking. If the target protein has been previously characterised, then information such as subcellular location, function, molecular weight and pI can be used for the design of a purification strategy. However, if the target protein is uncharacterised or little information regarding its characteristics is available, a generic purification protocol can be employed that is optimised as additional characteristics of the target protein are determined during subsequent purification steps. Herein, we describe the protocol for the purification and detection of a poly-γ-D-glutamic acid (PDGA) depolymerase from a consortium culture of two Gram-negative bacteria using a combination of chromatography, ...
[摘要] 从蛋白质的复杂混合物中纯化靶蛋白是一项具有挑战性的任务。 如果靶蛋白以前已被表征，则诸如亚细胞定位，功能，分子量和pI的信息可用于设计纯化策略。 然而，如果目标蛋白质是未表征的或者关于其特征的很少的信息可用，则可以使用通用的纯化方案，其随着在随后的纯化步骤期间测定靶标蛋白质的附加特征而被优化。 在这里，我们描述了从两个革兰氏阴性细菌的联合培养物使用色谱，2D电泳和酶谱的组合纯化和检测聚-γ-D-谷氨酸（PDGA）解聚酶的协议。
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| ChIP-Seq in Candida albicans
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Author:
Date:
2014-06-20
[Abstract] Systems biology approaches can be used to study the regulatory interactions occurring between many components of the biological system at the whole-genome level and decipher the circuitries implicated in the regulation of cellular processes, including those imparting virulence to opportunistic fungi. Candida albicans (C. albicans) is a leading human fungal pathogen. It undergoes morphological switching between a budding yeast form and an elongated multicellular hyphal form. This transition is required for C. albicans’ ability to cause disease and is regulated through highly interconnected regulatory interactions between transcription factors (TFs) and target genes. The chromatin immunoprecipitation (ChIP)-High-throughput sequencing (Seq) technology (ChIP-Seq) is ...
[摘要] 系统生物学方法可以用于研究在全基因组水平上发生在生物系统的许多组分之间的调节相互作用，并且解释涉及细胞过程调节的电路，包括赋予机会性真菌的毒力的电路。 白色念珠菌（ C。albicans ）是一种主要的人类真菌病原体。它经历萌芽酵母形式和细长多细胞菌丝形式之间的形态学转换。此转换是 C所必需的。白色念珠菌引起疾病的能力，并且通过转录因子（TF）和靶基因之间高度相互关联的调节相互作用来调节。染色质免疫沉淀（ChIP） - 高通量测序（Seq）技术（ChIP-Seq）是解码转录调节网络的强有力的方法。该方案针对从丝状结构C制备ChIP DNA进行了优化。白色念珠菌细胞，然后进行高通量测序以鉴定调节酵母对菌丝转变的TF的靶标。
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