| Quantification of Low Molecular Weight Thiols in Arabidopsis
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Author:
Date:
2016-01-05
[Abstract] Low-molecular-weight (LMW) thiols are a class of highly reactive compounds due to their thiol moiety. They play important roles in the maintenance of cellular redox homeostasis, detoxification, and development. Monobromobimane (mBBr) is weakly fluorescent but selectively reacts with thiols to yield highly fluorescent thioethers (mBSR) products, which is especially useful for the quantification of LMW thiols. The stable mBSR products can be separated by high-performance liquid chromatography (HPLC) equipped with a fluorescent detector. The main cellular LMW thiols are L-cysteine, gamma-glutamylcysteine, and glutathione (GSH). The following protocol describes the extraction and quantification of L-cysteine, gamma-glutamylcysteine, and glutathione from Arabidopsis tissues as ...
[摘要] 低分子量（LMW）硫醇是一类高反应性化合物，由于它们的硫醇部分。 它们在维持细胞氧化还原稳态，解毒和发育中起重要作用。 单溴溴烷（mBBr）是弱荧光的，但选择性地与硫醇反应产生高荧光硫醚（mBSR）产物，其对于定量LMW硫醇特别有用。 稳定的mBSR产物可以通过配备有荧光检测器的高效液相色谱（HPLC）分离。 主要的细胞LMW硫醇是L-半胱氨酸，γ-谷氨酰半胱氨酸和谷胱甘肽（GSH）。 以下方案描述了从所报道的拟南芥组织中提取和定量L-半胱氨酸，γ-谷氨酰半胱氨酸和谷胱甘肽（Xiang和Oliver，1998; Zhao等人， 2014; Wang ，，2015）。 如果HPLC系统或C18柱不同，可能需要进行修改。
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| Characterization of HBV Isolates from Patient Serum Samples and Cloning
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Author:
Date:
2015-12-20
[Abstract] Hepatitis B virus (HBV) mutants can lead to vaccine failure, diagnostic failure of HBV detection, increase viral replication and resistance to antiviral agents. To study the biological characteristics of these mutations may contribute to our knowledge on viral pathogenesis. Therefore, it is essential to isolate and characterize HBV strains from patients. Here we describe the experimental methods to isolate and clone HBV DNA from patient serum. The method will facilitate isolation and functional analysis of new HBV variants.
[摘要] 乙型肝炎病毒（HBV）突变体可导致疫苗失败，HBV检测的诊断失败，增加病毒复制和对抗病毒剂的抗性。 研究这些突变的生物学特性可能有助于我们对病毒发病机制的了解。 因此，必须从患者中分离和表征HBV病毒株。 在这里我们描述从患者血清中分离和克隆HBV DNA的实验方法。 该方法将促进新的HBV变体的分离和功能分析。
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