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    现刊
    Targeted Genome Editing of Virulent Phages Using CRISPR-Cas9
    使用CRISPR-Cas9对烈性噬菌体进行靶向基因组编辑
    作者:Marie-Laurence Lemay, Ariane C. Renaud, Geneviève M. Rousseau and Sylvain Moineau日期:01/05/2018,浏览量:219,Q&A: 0
    This protocol describes a straightforward method to generate specific mutations in the genome of strictly lytic phages. Briefly, a targeting CRISPR-Cas9 system and a repair template suited for homologous recombination are provided inside a bacterial ...
    Analysis of Direct Interaction between Viral DNA-binding Proteins by Protein Pull-down Co-immunoprecipitation Assay
    蛋白质pull-down免疫共沉淀实验分析病毒DNA结合蛋白质之间的直接相互作用
    作者:Ana Lechuga, Mónica Berjón-Otero, Margarita Salas and Modesto Redrejo-Rodríguez日期:01/05/2018,浏览量:221,Q&A: 0
    This protocol analyzes the direct interaction between two DNA-binding proteins by pull-down co-immunoprecipitation. One of the proteins is overexpressed in E. coli as HA-tagged recombinant protein and cell-free extracts are ...
    Visible Immunoprecipitation (VIP) Assay: a Simple and Versatile Method for Visual Detection of Protein-protein Interactions
    可视免疫沉淀(VIP)实验:一种简单通用的蛋白质 - 蛋白质相互作用可视化检测方法
    作者:Yohei Katoh, Kentaro Nakamura and Kazuhisa Nakayama日期:01/05/2018,浏览量:232,Q&A: 0
    The visible immunoprecipitation (VIP) assay is a convenient alternative to conventional co-immunoprecipitation (Katoh et al., 2015). By processing lysates from cells co-expressing GFP-fusion and RFP-fusion proteins for immunoprecipitation ...
    Bacterial Aggregation Assay in the Presence of Cyclic Lipopeptides
    环脂肽存在情况下细菌聚集实验
    作者:Pengyuan Xiu, Rui Liu, Dechao Zhang and Chaomin Sun日期:01/05/2018,浏览量:83,Q&A: 0
    Lipopeptides is an important class of biosurfactants having antimicrobial and anti-adhesive activity against pathogenic bacteria. These include surfactin, fengycin, iturin, bacillomycin, mycosubtilin, lichenysin, and pumilacidin (Arima et al. ...
    Easy and Efficient Permeabilization of Cyanobacteria for in vivo Enzyme Assays Using B-PER
    使用B-PER简单有效地透化蓝藻以进行体内酶分析
    作者:Simon Matthé Erstad and Yumiko Sakuragi日期:01/05/2018,浏览量:172,Q&A: 0
    Cyanobacteria are photosynthetic bacteria that thrive in diverse ecosystems and play major roles in the global carbon cycle. The abilities of cyanobacteria to fix atmospheric CO2 and to allocate the fixed carbons to chemicals and biofuels ...
    Targeted Genome Editing of Virulent Phages Using CRISPR-Cas9
    使用CRISPR-Cas9对烈性噬菌体进行靶向基因组编辑
    作者:Marie-Laurence Lemay, Ariane C. Renaud, Geneviève M. Rousseau and Sylvain Moineau日期:01/05/2018,浏览量:219,Q&A: 0
    [Abstract] This protocol describes a straightforward method to generate specific mutations in the genome of strictly lytic phages. Briefly, a targeting CRISPR-Cas9 system and a repair template suited for homologous recombination are provided inside a bacterial host, here the Gram-positive model Lactococcus lactis MG1363. The CRISPR-Cas9 system is ...
    Analysis of Direct Interaction between Viral DNA-binding Proteins by Protein Pull-down Co-immunoprecipitation Assay
    蛋白质pull-down免疫共沉淀实验分析病毒DNA结合蛋白质之间的直接相互作用
    作者:Ana Lechuga, Mónica Berjón-Otero, Margarita Salas and Modesto Redrejo-Rodríguez日期:01/05/2018,浏览量:221,Q&A: 0
    [Abstract] This protocol analyzes the direct interaction between two DNA-binding proteins by pull-down co-immunoprecipitation. One of the proteins is overexpressed in E. coli as HA-tagged recombinant protein and cell-free extracts are immunoprecipitated in HA-affinity resin. Cell extracts are treated with nuclease to degrade DNA and RNA, which rules ...
    Visible Immunoprecipitation (VIP) Assay: a Simple and Versatile Method for Visual Detection of Protein-protein Interactions
    可视免疫沉淀(VIP)实验:一种简单通用的蛋白质 - 蛋白质相互作用可视化检测方法
    作者:Yohei Katoh, Kentaro Nakamura and Kazuhisa Nakayama日期:01/05/2018,浏览量:232,Q&A: 0
    [Abstract] The visible immunoprecipitation (VIP) assay is a convenient alternative to conventional co-immunoprecipitation (Katoh et al., 2015). By processing lysates from cells co-expressing GFP-fusion and RFP-fusion proteins for immunoprecipitation with GST-tagged anti-GFP Nanobody and glutathione-Sepharose beads, protein-protein interactions can ...
    Bacterial Aggregation Assay in the Presence of Cyclic Lipopeptides
    环脂肽存在情况下细菌聚集实验
    作者:Pengyuan Xiu, Rui Liu, Dechao Zhang and Chaomin Sun日期:01/05/2018,浏览量:83,Q&A: 0
    [Abstract] Lipopeptides is an important class of biosurfactants having antimicrobial and anti-adhesive activity against pathogenic bacteria. These include surfactin, fengycin, iturin, bacillomycin, mycosubtilin, lichenysin, and pumilacidin (Arima et al., 1968; Naruse et al., 1990; Yakimov et al., 1995; Steller and Vater, 2000; ...
    Easy and Efficient Permeabilization of Cyanobacteria for in vivo Enzyme Assays Using B-PER
    使用B-PER简单有效地透化蓝藻以进行体内酶分析
    作者:Simon Matthé Erstad and Yumiko Sakuragi日期:01/05/2018,浏览量:172,Q&A: 0
    [Abstract] Cyanobacteria are photosynthetic bacteria that thrive in diverse ecosystems and play major roles in the global carbon cycle. The abilities of cyanobacteria to fix atmospheric CO2 and to allocate the fixed carbons to chemicals and biofuels have attracted growing attentions as sustainable microbial cell factories. A better understanding ...
    Infection of Soybean Plants with the Insect Bacterial Symbiont Burkholderia gladioli and Evaluation of Plant Fitness
    用昆虫细菌共生体唐菖蒲伯克霍尔德菌感染大豆植株及植物适应性评价
    作者:Paul Gaube, Martin Kaltenpoth and Laura V. Flórez日期:12/20/2017,浏览量:399,Q&A: 0
    [Abstract] To investigate the establishment and consequences of host-microbe interactions, it is important to develop controlled infection assays suitable for each system, as well as appropriate methods to evaluate successful infection and its associated effects. Here, we describe a procedure for bacterial inoculation of soybean plants, followed by the ...
    Determination of Mn Concentrations in Synechocystis sp. PCC6803 Using ICP-MS
    采用ICP-MS测定集胞藻PCC6803中Mn的浓度
    作者:Fabian Brandenburg, Hanan Schoffman, Nir Keren and Marion Eisenhut日期:12/05/2017,浏览量:404,Q&A: 0
    [Abstract] Manganese (Mn) is an essential micronutrient for all photoautotrophic organisms. Two distinct pools of Mn have been identified in the cyanobacterium Synechocystis sp. PCC 6803 (Synechocystis), with 80% of the Mn residing in the periplasm and 20% in cytoplasm and thylakoid lumen (Keren et al., 2002). In this protocol, we ...
    Quantification of Bacterial Polyhydroxybutyrate Content by Flow Cytometry
    流式细胞术定量分析细菌聚羟基丁酸酯含量
    作者:Antonio Lagares (Jr.) and Claudio Valverde日期:12/05/2017,浏览量:319,Q&A: 0
    [Abstract] We describe here a detailed protocol for the quantification of the intracellular content of polyhydroxybutyrate (PHB) in a population of bacterial cells by flow cytometry, which is based on a consensus of several previously reported works.
    Stationary-phase Mutagenesis Soft-agar Overlay Assays in Bacillus subtilis
    枯草芽孢杆菌的软琼脂糖覆盖固相诱变
    [Abstract] Elucidating how a population of non-growing bacteria generates mutations improves our understanding of phenomena like antibiotic resistance, bacterial pathogenesis, genetic diversity and evolution. To evaluate mutations that occur in nutritionally stressed non-growing bacteria, we have employed the strain B. subtilis YB955, which measures ...
    Design and Direct Assembly of Synthesized Uracil-containing Non-clonal DNA Fragments into Vectors by USERTM Cloning
    设计并通过USERTM克隆直接将合成的含尿嘧啶非克隆DNA片段装配到载体中
    [Abstract] This protocol describes how to order and directly assemble uracil-containing non-clonal DNA fragments by uracil excision based cloning (USER cloning). The protocol was generated with the goal of making synthesized non-clonal DNA fragments directly compatible with USERTM cloning. The protocol is highly efficient and would be compatible ...