微生物学

分类

    现刊
    A New Tool for the Flexible Genetic Manipulation of Geobacillus kaustophilus
    一种对嗜热土杆菌进行灵活遗传操作的新工具
    作者:Ryotaro Amatsu, Kotaro Mori, Shu Ishikawa, Wilfried J. J. Meijer and Ken-ichi Yoshida日期:09/05/2022,浏览量:601,Q&A: 0
    [Abstract]

    Geobacillus kaustophilus, a thermophilic Gram-positive bacterium, is an attractive host for the development of high-temperature bioprocesses. However, its reluctance against genetic manipulation by standard methodologies hampers its exploitation. Here, we describe a simple methodology in which an artificial DNA segment on the chromosome of

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    A Modified Fluctuation Assay with a CAN1 Reporter in Yeast
    酵母中CAN1报告基因的改良波动测定
    作者:Pengyao Jiang, Anja R. Ollodart and Maitreya J. Dunham日期:06/05/2022,浏览量:1017,Q&A: 0
    [Abstract]

    Understanding the generation of mutations is fundamental to understanding evolution and genetic disease; however, the rarity of such events makes experimentally identifying them difficult. Mutation accumulation (MA) methods have been widely used. MA lines require serial bottlenecks to fix de novo mutations, followed by whole-genome sequencing.

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    A Molecular Cloning and Sanger Sequencing-based Protocol for Detecting Site-specific DNA Methylation
    一种基于分子克隆和Sanger测序的位点特异性DNA甲基化检测方案
    作者:Wei Guo, Anthony Cannon and Damon Lisch日期:05/05/2022,浏览量:1460,Q&A: 1
    [Abstract]

    DNA methylation is a conserved chemical modification, by which methyl groups are added to the cytosine of DNA molecules. Methylation can influence gene expression without changing the sequence of a particular gene. This epigenetic effect is an intriguing phenomenon that has puzzled biologists for years. By probing the temporal and spatial patterns

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    Phytophthora sojae Transformation Based on the CRISPR/Cas9 System
    基于CRISPR/Cas9系统的大豆疫霉转化
    作者:Jingting Cao, Min Qiu, Wenwu Ye and Yuanchao Wang日期:03/20/2022,浏览量:1374,Q&A: 0
    [Abstract]

    Phytophthora sojae is a model species for the study of plant pathogenic oomycetes. The initial research on gene function using Phytophthora was mainly based on gene silencing technology. Recently, the CRISPR/Cas9-mediated genome editing technology was successfully established in P. sojae and widely used in oomycetes. In this protocol, we describe

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    RNA-mediated in vivo Directed Evolution in Yeast
    RNA介导的酵母体内定向进化
    作者:Emil D. Jensen and Michael K. Jensen日期:03/05/2022,浏览量:2027,Q&A: 0
    [Abstract]

    Directed evolution is a powerful approach to obtain genetically-encoded sought-for traits. Compared to the prolonged adaptation regimes to mutations occurring under natural selection, directed evolution unlocks rapid screening and selection of mutants with improved traits from vast mutated sequence spaces. Many systems have been developed to

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    An in vitro Assay of mRNA 3’ end Using the E. coli Cell-free Expression System
    使用大肠杆菌无细胞表达系统对 mRNA 3' 端进行体外测定
    作者:Monford Paul Abishek N and Heon M. Lim日期:02/20/2022,浏览量:1747,Q&A: 0
    [Abstract]

    At the end of about 80% of the operon in Escherichia coli, translation termination decouples transcription, leading to Rho-dependent transcription termination (RDT). However, no in vitro or in vivo assay system has proven to be good enough to see the 3’ end of the mRNA generated by RDT. Here, we present a cell-free assay system that

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    Selection of Vaccinia Virus Recombinants Using CRISPR/Cas9
    使用 CRISPR/Cas9 筛选牛痘病毒重组体
    作者:Anjali Gowripalan, Stewart A. Smith and David C. Tscharke日期:12/20/2021,浏览量:1698,Q&A: 0
    [Abstract]

    The engineering of poxvirus genomes is fundamental to primary and applied virology research. Indeed, recombinant poxviruses form the basis for many novel vaccines and virotherapies but producing and purifying these viruses can be arduous. In recent years, CRISPR/Cas9 has become the favoured approach for genome manipulation due to its speed and

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    A Nucleocapsid-based Transcomplementation Cell Culture System of SARS-CoV-2 to Recapitulate the Complete Viral Life Cycle
    基于核衣壳的 SARS-CoV-2 转互补细胞培养系统,重现完整的病毒生命周期
    作者:Yanying Yu, Xiaohui Ju and Qiang Ding日期:11/05/2021,浏览量:1951,Q&A: 0
    [Abstract]

    The ongoing COVID-19 pandemic is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). As this virus is classified as a biosafety level-3 (BSL-3) agent, the development of countermeasures and basic research methods is logistically difficult. Recently, using reverse genetics, we developed a BSL-2 cell culture system for production

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    A Rapid Induction Overexpression System for the Fission Yeast Schizosaccharomyces pombe
    裂解酵母裂殖酵母的快速诱导过表达系统
    作者:Angad Garg日期:10/20/2021,浏览量:1219,Q&A: 0
    [Abstract]

    The fission yeast Schizosaccharomyces pombe is an excellent genetically tractable model organism used in the study of conserved eukaryotic cellular biology. One genetic tool in the assessment of gene function is the in vivo overexpression of proteins. Existing overexpression tools have limitations of induction kinetics, dynamic range, and/or

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    Quantification of RuBisCO Expression and Photosynthetic Oxygen Evolution in Cyanobacteria
    蓝藻RuBisCO表达和光合氧释放的定量研究
    作者:Mateusz Kędzior and Betul Kacar日期:10/20/2021,浏览量:1474,Q&A: 0
    [Abstract]

    Phototrophic microorganisms are frequently engineered to regulate the expression and the activity of targeted enzymes of interest for specific biotechnological and agricultural applications. This protocol describes a method to evaluate the expression of RuBisCO (ribulose 1,5-bisphosphate carboxylase/oxygenase) in the model cyanobacterium

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