| Catalase Activity Assay in Candida glabrata
|
|
Author:
Date:
2014-03-20
[Abstract] Commensal and pathogenic fungi are exposed to hydrogen peroxide (H2O2) produced by macrophages of the host. Pathogenic fungi counteract the harmful effects of H2O2 with the enzyme catalase (EC 1.11.1.6), which decomposes two molecules of H2O2 to two molecules of H2O and O2. Contribution of antioxidant systems on fungal virulence is actively studied. Measurement of catalase activity can contribute to the elucidation of the factors that influence the regulation of this pivotal enzyme. Here we describe a simple spectrophotometric method in which the activity of catalase is measured in total yeast extracts. Decomposition of H2O2 by the yeast extract is followed by the decrease in ...
[摘要] 共生和致病真菌暴露于由宿主的巨噬细胞产生的过氧化氢（H 2 O 2 O 2）。 致病真菌抵消了H 2 O 2对于过氧化氢酶（EC 1.11.1.6）的有害影响，所述过氧化氢酶分解两个分子的H 2 O 2 - O 2至两个H 2 O和O 2分子。 积极研究抗氧化系统对真菌毒力的贡献。 过氧化氢酶活性的测量可有助于阐明影响这种关键酶的调节的因素。 在这里我们描述一个简单的分光光度法，其中过氧化氢酶的活性在总酵母提取物中测量。 通过酵母提取物的H 2 O 2 O 2分解后，在240nm处的吸光度降低。 吸光度随时间的差异（ΔA240）被推断为过氧化氢酶活性的量度。
|
|
|
| GST-tagged Yeast Protein Purification
|
|
Author:
Date:
2011-10-05
[Abstract] Glutation S-transferase (GST) tagging is the most commonly used purification strategy for recombinant protein. It was developed with the goal of preserving the enzymatic activity by utilizing gentle elution condition of the target protein from purification matrix (Poon and Hunt., 1994). The method described here can be applied from single protein to proteome scale purification of recombinant protein from yeast (Zhu et al., 2000; Zhu et al., 2001).
[摘要] Gligation S转移酶（GST）标签是重组蛋白最常用的纯化策略。 它的开发目的是通过利用来自纯化基质的目标蛋白的温和洗脱条件来保持酶活性（Poon和Hunt，1994）。 本文所述的方法可以从单一蛋白质应用于来自酵母的重组蛋白质的蛋白质组规模纯化（Zhu et al。，2000; Zhu et al。，2001）。
|
|
|
| Yeast Transcription Factor Chromatin Immunoprecipitation
|
|
Author:
Date:
2011-07-05
[Abstract] This ChIP protocol was developed and improved over the years by various researchers in the Snyder lab, Stanford University, especially Anthony Borneman and Christopher Yellman. I have used this method to successfully map the genome-wide binding of transcription factors Ste12. The ChIPed DNA is suitable for downstream analysis using PCR, microarray or sequencing.
[摘要] 这个ChIP协议是由斯坦福大学斯奈德实验室，特别是安东尼博恩曼和克里斯托弗·耶尔曼的研究人员多年来开发和改进的。 我已经使用这种方法成功映射转录因子Ste12的全基因组结合。 ChIPed DNA适用于使用PCR，微阵列或测序的下游分析。
|
|
|