微生物学

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    现刊
    In vitro Nitrate Reductase Activity Assay of Mycolicibacterium smegmatis Crude Extract
    耻垢分枝杆菌粗提物硝酸还原酶活性的体外测定
    作者:Wei Tan, Zhi-Hui Shao and Guo-Ping Zhao日期:07/20/2021,浏览量:640,Q&A: 0
    [Abstract]

    Nitrate is one of the major inorganic nitrogen sources for microorganisms. Many bacterial and archaeal lineages can express cytoplasmic assimilatory nitrate reductase (NAS), which catalyzes the rate-limiting reduction of nitrate to nitrite in the nitrate assimilation pathway. Here, we present a detailed protocol for measuring in vitro nitrate

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    In vitro Glutamylation Inhibition of Ubiquitin Modification and Phosphoribosyl-Ubiquitin Ligation Mediated by Legionella pneumophila Effectors
    嗜肺军团菌介导的泛素修饰和磷酸核糖泛素连接的体外谷氨酰化抑制
    作者:Alan G. Sulpizio, Marena E. Minelli and Yuxin Mao日期:11/05/2020,浏览量:1164,Q&A: 0
    [Abstract]

    Glutamylation is a posttranslational modification where the amino group of a free glutamate amino acid is conjugated to the carboxyl group of a glutamate side chain within a target protein. SidJ is a Legionella kinase-like protein that has recently been identified to perform protein polyglutamylation of the Legionella SdeA Phosphoribosyl-Ubiquitin

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    A Sensitive Coupled Enzyme Assay for Measuring Kinase and ATPase Kinetics Using ADP-Specific Hexokinase
    使用ADP特异性己糖激酶测量激酶和ATPase动力学的灵敏耦合酶活测定
    作者:Ciaran R. McFarlane and James W. Murray日期:05/05/2020,浏览量:3379,Q&A: 0
    [Abstract] Kinases and ATPases perform essential biological functions in metabolism and regulation. Activity of these enzymes is commonly measured by coupling ATP consumption to the synthesis of a detectable product. For most assay systems the ATP concentration during the reaction is unknown, compromising the precision of the assay.

    Using the ...
    Assessment of Metacaspase Activity in Phytoplankton
    浮游植物中半胱天冬酶活性检测
    作者:Dina Spungin and Ilana Berman-Frank日期:08/20/2019,浏览量:2639,Q&A: 0
    [Abstract] Programmed cell death (PCD) is an irreversible, genetically-controlled form of cell suicide in which an endogenous biochemical pathway leads to morphological changes and ultimately, cellular demise. PCD is accompanied by de-novo protein synthesis of a family of proteases-"caspases" that are often used as a diagnostic marker of PCD. ...
    Kinetic Characterization of the Shigella Type Three Secretion System ATPase Spa47 Using α-32P ATP
    利用α-32P ATP研究志贺氏菌的III型分泌系统ATP酶Spa47的动力学表征
    作者:Heather B. Case and Nicholas E. Dickenson日期:11/05/2018,浏览量:3215,Q&A: 0
    [Abstract] ATPases represent a diverse class of enzymes that utilize ATP hydrolysis to support critical biological functions such as driving ion pumps, providing mechanical work, unfolding/folding proteins, and supporting otherwise thermodynamically unfavorable chemical reactions. We have recently shown that the Shigella protein Spa47 is an ATPase ...
    Detection of Catalase Activity by Polyacrylamide Gel Electrophoresis (PAGE) in Cell Extracts from Pseudomonas aeruginosa
    聚丙烯酰胺凝胶电泳(PAGE)检测铜绿假单胞菌细胞提取物中的过氧化氢酶活性
    作者:Magdalena Pezzoni, Ramón A. Pizarro and Cristina S. Costa日期:06/05/2018,浏览量:6437,Q&A: 0
    [Abstract] Bacteria in nature and as pathogens commonly face oxidative stress which causes damage to proteins, lipids and DNA. This damage is produced by the action of reactive oxygen species (ROS) such as hydrogen peroxide (H2O2), singlet oxygen, superoxide anion and hydroxyl radical. ROS are generated by antimicrobials, environmental ...
    In vitro Analysis of Ubiquitin-like Protein Modification in Archaea
    古生菌中泛素样蛋白修饰的体外分析
    作者:Xian Fu, Zachary Adams and Julie A. Maupin-Furlow日期:05/20/2018,浏览量:4613,Q&A: 0
    [Abstract] The ubiquitin-like (Ubl) protein is widely distributed in Archaea and involved in many cellular pathways. A well-established method to reconstitute archaeal Ubl protein conjugation in vitro is important to better understand the process of archaeal Ubl protein modification. This protocol describes the in vitro reconstitution of ...
    Heterologous Expression and Purification of the CRISPR-Cas12a/Cpf1 Protein
    CRISPR-Cas12a/Cpf1的异源表达和纯化
    作者:Prarthana Mohanraju, John van der Oost, Martin Jinek and Daan C. Swarts日期:05/05/2018,浏览量:11756,Q&A: 3
    [Abstract] This protocol provides step by step instructions (Figure 1) for heterologous expression of Francisella novicida Cas12a (previously known as Cpf1) in Escherichia coli. It additionally includes a protocol for high-purity purification and briefly describes how activity assays can be performed. These protocols can also be used for ...
    Superoxide Dismutase (SOD) and Catalase (CAT) Activity Assay Protocols for Caenorhabditis elegans
    秀丽隐杆线虫中超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性测定实验方案
    作者:Jing Zhang, Rui Chen, Zhenyang Yu and Lili Xue日期:08/20/2017,浏览量:31268,Q&A: 0
    [Abstract] Assays for superoxide dismutase (SOD) and catalase (CAT) activities are widely employed to indicate antioxidant responses underlying the toxic effects of test chemicals. Yet, earlier studies mainly described the procedures as performed according to manufacturer’s instructions without modifications that are specific to any organisms. The present ...
    Liposome Disruption Assay to Examine Lytic Properties of Biomolecules
    采用脂质体破碎测定法检测生物分子的裂解性质
    作者:John R. Jimah, Paul H. Schlesinger and Niraj H. Tolia日期:08/05/2017,浏览量:10283,Q&A: 0
    [Abstract] Proteins may have three dimensional structural or amino acid features that suggest a role in targeting and disrupting lipids within cell membranes. It is often necessary to experimentally investigate if these proteins and biomolecules are able to disrupt membranes in order to conclusively characterize the function of these biomolecules. Here, we ...