微生物学

分类

    现刊
    Construction of a Highly Diverse mRNA Library for in vitro Selection of Monobodies
    体外选择单体的高度多样性mRNA文库的构建
    [Abstract]

    Recently, we developed transcription/translation coupled with the association of puromycin linker (TRAP) display as a quick in vitro selection method to obtain antibody-like proteins. For the in vitro selection, it is important to prepare mRNA libraries among which the diversity is high. Here, we describe a method for the preparation of monobody

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    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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    Assay for Assessing Mucin Binding to Bacteria and Bacterial Proteins
    评估粘蛋白与细菌和细菌蛋白结合的实验
    作者:Lubov S. Grigoryeva, Saima Rehman, Richard C. White, James A. Garnett and Nicholas P. Cianciotto日期:03/05/2021,浏览量:1584,Q&A: 0
    [Abstract]

    Legionella pneumophila, a Gram-negative bacterium and the causative agent of Legionnaires’ disease, exports over 300 effector proteins/virulence factors, through its type II (T2SS) and type IV secretion systems (T4SS). One such T2SS virulence factor, ChiA, not only functions as a chitinase, but also as a novel mucinase, which we believe aids

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    Preparation of Bacterial Outer Membrane Vesicles for Characterisation of Periplasmic Proteins in Their Native Environment
    细菌外膜囊泡的制备及其原生环境中周质蛋白的表征
    作者:Johannes Thoma and Björn M. Burmann日期:12/20/2020,浏览量:1710,Q&A: 0
    [Abstract]

    Bacterial outer membrane vesicles (OMVs) are naturally formed by budding from the outer membrane of Gram-negative bacteria. OMVs consist of a lipid bilayer identical in composition to the original outer membrane and contain periplasmic content within their lumen. Enriched with specific envelope proteins, OMVs make for an excellent native-like

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    Expression and Purification of Recombinant Skd3 (Human ClpB) Protein and Tobacco Etch Virus (TEV) Protease from Escherichia coli
    重组Skd3(人ClpB)蛋白和烟草蚀刻病毒(TEV)蛋白酶在大肠杆菌中的表达与纯化
    作者:Ryan R. Cupo and James Shorter日期:12/05/2020,浏览量:1853,Q&A: 0
    [Abstract]

    Skd3 (encoded by human CLPB) is a mitochondrial AAA+ protein comprised of an N-terminal ankyrin-repeat domain and a C-terminal HCLR-clade nucleotide-binding domain. The function of Skd3 has long remained unknown due to challenges in purifying the protein to high quality and near homogeneity. Recently we described Skd3 as a human mitochondrial

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    Combining Gel Retardation and Footprinting to Determine Protein-DNA Interactions of Specific and/or Less Stable Complexes
    结合凝胶阻滞和足印法测定特异性和/或不稳定复合物的蛋白质-DNA相互作用
    作者:Meng-Lun Hsieh, Alice Boulanger, Leslie G. Knipling and Deborah M. Hinton日期:12/05/2020,浏览量:1080,Q&A: 0
    [Abstract]

    DNA footprinting is a classic technique to investigate protein-DNA interactions. However, traditional footprinting protocols can be unsuccessful or difficult to interpret if the binding of the protein to the DNA is weak, the protein has a fast off-rate, or if several different protein-DNA complexes are formed. Our protocol differs from traditional

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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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    Radioactive Assay of in vitro Glutamylation Activity of the Legionella pneumophila Effector Protein SidJ
    嗜肺军团菌效应蛋白SidJ体外谷氨酰化活性的放射性测定
    作者:Alan G. Sulpizio, Jung-Ho Shin, Marena E. Minelli and Yuxin Mao日期:10/05/2020,浏览量:1510,Q&A: 0
    [Abstract] The Legionella effector protein SidJ has recently been identified to perform polyglutamylation on another Legionella effector, SdeA, ablating SdeA’s activity. SidJ is a kinase-like protein that requires the small eukaryotic protein calmodulin to perform glutamylation. Glutamylation is a relatively uncommon type of ...
    A SsrA/NIa-based Strategy for Post-Translational Regulation of Protein Levels in Gram-negative Bacteria
    基于SsrA/NIa的革兰阴性菌蛋白质水平翻译后调控策略
    作者:Gonzalo Durante-Rodríguez, Belén Calles, Víctor de Lorenzo and Pablo I. Nikel日期:07/20/2020,浏览量:1672,Q&A: 0
    [Abstract] Strategies to control the levels of key enzymes of bacterial metabolism are commonly based on the manipulation of gene of interest within the target pathway. The development of new protocols towards the manipulation of biochemical processes is still a major challenge in the field of metabolic engineering. On this background, the FENIX (functional ...
    Identification of Buffer Conditions for Optimal Thermostability and Solubility of Herpesviral Protein UL37 Using the Thermofluor Assay
    用热荧光法鉴定疱疹病毒蛋白UL37的最佳热稳定性和溶解度的缓冲液条件
    作者:Andrea L. Koenigsberg, Jared D. Pitts and Ekaterina E. Heldwein日期:06/20/2020,浏览量:1658,Q&A: 0
    [Abstract] Structural and biochemical studies of proteins require high amounts of stable, purified proteins. Protein stability often depends on the buffer composition, which includes pH and concentration of salts or other solutes such as glycerol, hence an efficient method for identifying optimal buffer conditions for stability would minimize time and ...