| Catalase Activity Assay in Candida glabrata
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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 ...
[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。
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| GST-tagged Yeast Protein Purification
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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] 谷胱甘肽S 转移酶（GST）标记是最常用的重组蛋白的纯化方法。通过利用温和的洗脱条件以保护酶的活性，使靶蛋白高纯化(1)。这里介绍的方法可以适用于从单一的蛋白质，以及蛋白质组大规模纯化重组蛋白酵母（2,3）。
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| Yeast Transcription Factor Chromatin Immunoprecipitation
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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] 本篇 ChIP 的方法是Snyder实验室多年来经多位学者改进优化而来，特别是 Anthony Borneman 和 Christopher Yellman。我应用此方法已成功定位了转录因子Ste12在基因组范围内的结合。ChIP得到的DNA可通过PCR、芯片及测序进行下游的分析。
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