分子生物学

分类

    现刊
    A Sensitive and Specific PCR-based Assay to Quantify Hepatitis B Virus Covalently Closed Circular (ccc) DNA while Preserving Cellular DNA
    一种在保留细胞DNA的同时定量检测乙型肝炎病毒共价闭合环状(ccc)DNA的灵敏、特异PCR方法
    作者:Benno Zehnder, Stephan Urban and Thomas Tu日期:04/20/2021,浏览量:0,Q&A: 0

    Hepatitis B virus (HBV) is the major cause of liver diseases and liver cancer worldwide. After infecting hepatocytes, the virus establishes a stable episome (covalently closed circular DNA, or cccDNA) that serves as the template for all viral

    ...
    Optimized Recombinant Production of Secreted Proteins Using Human Embryonic Kidney (HEK293) Cells Grown in Suspension
    利用悬浮培养的人胚胎肾(HEK293)细胞优化分泌蛋白的重组生产

    Recombinant proteins are an essential milestone for a plethora of different applications ranging from pharmaceutical to clinical, and mammalian cell lines are among the currently preferred systems to obtain large amounts of proteins of interest due

    ...
    Optimized Recombinant Production of Secreted Proteins Using Human Embryonic Kidney (HEK293) Cells Grown in Suspension
    利用悬浮培养的人胚胎肾(HEK293)细胞优化分泌蛋白的重组生产
    [Abstract]

    Recombinant proteins are an essential milestone for a plethora of different applications ranging from pharmaceutical to clinical, and mammalian cell lines are among the currently preferred systems to obtain large amounts of proteins of interest due to their high level of post-translational modification and manageable large-scale production. In

    ...
    A Sensitive and Specific PCR-based Assay to Quantify Hepatitis B Virus Covalently Closed Circular (ccc) DNA while Preserving Cellular DNA
    一种在保留细胞DNA的同时定量检测乙型肝炎病毒共价闭合环状(ccc)DNA的灵敏、特异PCR方法
    作者:Benno Zehnder, Stephan Urban and Thomas Tu日期:04/20/2021,浏览量:0,Q&A: 0
    [Abstract]

    Hepatitis B virus (HBV) is the major cause of liver diseases and liver cancer worldwide. After infecting hepatocytes, the virus establishes a stable episome (covalently closed circular DNA, or cccDNA) that serves as the template for all viral transcripts. Specific and accurate quantification of cccDNA is difficult because infected cells contain

    ...
    Reconstitution of Chromatin by Stepwise Salt Dialysis
    逐步盐透析重组染色质
    作者:Grisel Cruz-Becerra and James T. Kadonaga日期:04/05/2021,浏览量:192,Q&A: 0
    [Abstract]

    Chromatin, rather than plain DNA, is the natural substrate of the molecular machines that mediate DNA-directed processes in the nucleus. Chromatin can be reconstituted in vitro by using different methodologies. The salt dialysis method yields chromatin that consists of purified histones and DNA. This biochemically pure chromatin is well-suited for

    ...
    A Novel Method to Construct Binary CRISPR Vectors for Plant Transformation by Single Round of PCR Amplification
    一种通过单轮PCR扩增构建用于植物转化的二元CRISPR载体的新方法
    作者:Kang Li, Yuhui Wang and Chuanying Fang日期:04/05/2021,浏览量:407,Q&A: 0
    [Abstract]

    CRISPR/Cas9 is an established and flexible tool for genome editing. However, most methods used to generate expression clones for the CRISPR/Cas9 are time-consuming. Hence, we have developed a one-step protocol to introduce sgRNA expression cassette(s) directly into binary vectors (Liu et al., 2020). In this approach, we have optimized the

    ...
    In vitro Reconstitution Assays of Arabidopsis 20S Proteasome
    拟南芥20S蛋白酶体的体外重组分析
    作者:Yanjun Li, Di Sun, Xingxing Yan, Zhiye Wang and Xiuren Zhang日期:04/05/2021,浏览量:203,Q&A: 0
    [Abstract]

    The majority of cellular proteins are degraded by the 26S proteasome in eukaryotes. However, intrinsically disordered proteins (IDPs), which contain large portions of unstructured regions and are inherently unstable, are degraded via the ubiquitin-independent 20S proteasome. Emerging evidence indicates that plant IDP homeostasis may also be

    ...
    Colorimetric RT-LAMP and LAMP-sequencing for Detecting SARS-CoV-2 RNA in Clinical Samples
    比色RT-LAMP和LAMP测序检测临床样本中SARS-CoV-2 RNA
    作者:Konrad Herbst, Matthias Meurer, Daniel Kirrmaier, Simon Anders, Michael Knop and Viet Loan Dao Thi日期:03/20/2021,浏览量:777,Q&A: 0
    [Abstract]

    During pandemics, such as the one caused by SARS-CoV-2 coronavirus, simple methods to rapidly test large numbers of people are needed. As a faster and less resource-demanding alternative to detect viral RNA by conventional qPCR, we used reverse transcription loop-mediated isothermal amplification (RT-LAMP). We previously established colorimetric

    ...
    Real-time Base Excision Repair Assay to Measure the Activity of the 8-oxoguanine DNA Glycosylase 1 in Isolated Mitochondria of Human Skin Fibroblasts
    实时碱基切除修复法测定人皮肤纤维细胞的分离线粒体中8-氧鸟嘌呤DNA糖基化酶1的活性
    作者:Daniel Schniertshauer, Daniel Gebhard and Jörg Bergemann日期:03/20/2021,浏览量:530,Q&A: 0
    [Abstract]

    7,8-dihydro-8-oxoguanine (8-oxoG) is one of the most common and mutagenic oxidative DNA damages induced by reactive oxygen species (ROS). Since ROS is mainly produced in the inner membranes of the mitochondria, these organelles and especially the mitochondrial DNA (mtDNA) contained therein are particularly affected by this damage. Insufficient

    ...
    Ligand and Carbohydrate Engagement (LACE) Assay and Fluorescence Quantification on Murine Neural Tissue
    小鼠神经组织中配体和碳水化合物结合(LACE)实验和荧光定量测定
    作者:James M. Clegg and Thomas Pratt日期:03/20/2021,浏览量:425,Q&A: 0
    [Abstract]

    The interaction between cell surface heparan sulphate and diffusible ligands such as FGFs is of vital importance for downstream signaling, however, there are few techniques that can be used to investigate this binding event. The ligand and carbohydrate engagement (LACE) assay is a powerful tool which can be used to probe the molecular interaction

    ...
    Transcardiac Perfusion of the Mouse for Brain Tissue Dissection and Fixation
    小鼠心脏灌流、脑组织解剖与固定
    作者:Jinyun Wu, Yuqi Cai, Xiaoyang Wu, Yue Ying, Yilin Tai and Miao He日期:03/05/2021,浏览量:829,Q&A: 0
    [Abstract] Transcardiac perfusion with saline followed by 4% paraformaldehyde (PFA) is widely used to clear blood and preserve brain for immunostaining or in situ hybridization. PFA breaks into formaldehyde in solution, which cross-link protein and DNA molecules to preserve tissue and cell structure. Here we provide a step by step guide for ...
    Defined Mutant Library Sequencing (DML-Seq) for Identification of Conditional Essential Genes
    确定突变体文库测序(DML-Seq)用于鉴定的条件必需基因
    作者:Shuai Shao, Lifan Wei, Feng Xia, Yuanxing Zhang and Qiyao Wang日期:03/05/2021,浏览量:437,Q&A: 0
    [Abstract]

    Transposon insertion sequencing (TIS) is an emerging technique which utilizes a massive transposon mutant library to screen specific phenotype and determine the conditional essential genetic requirements for bacterial fitness under distinct conditions combined with high-throughput parallel sequencing technology. Compared with a massive mutant

    ...