生物化学

分类

    现刊
    Fluorescent Polysome Profiling in Caenorhabditis elegans
    秀丽隐杆线虫荧光多聚体分析
    作者:Dan Shaffer and Jarod A Rollins日期:09/05/2020,浏览量:545,Q&A: 0
    An important but often overlooked aspect of gene regulation occurs at the level of protein translation. Many genes are regulated not only by transcription but by their propensity to be recruited to actively translating ribosomes (polysomes). ...
    Optogenetic Tuning of Protein-protein Binding in Bilayers Using LOVTRAP
    LOVTRAP对双层膜蛋白质结合的光遗传学调控
    作者:Doug Tischer and Orion D. Weiner日期:09/05/2020,浏览量:660,Q&A: 0
    Modern microscopy methods are powerful tools for studying live cell signaling and biochemical reactions, enabling us to observe when and where these reactions take place from the level of a cell down to single molecules. With microscopy, each cell ...
    Fluorescent Polysome Profiling in Caenorhabditis elegans
    秀丽隐杆线虫荧光多聚体分析
    作者:Dan Shaffer and Jarod A Rollins日期:09/05/2020,浏览量:545,Q&A: 0
    [Abstract] An important but often overlooked aspect of gene regulation occurs at the level of protein translation. Many genes are regulated not only by transcription but by their propensity to be recruited to actively translating ribosomes (polysomes). Polysome profiling allows for the separation of unbound 40S and 60S subunits, 80S monosomes, and actively ...
    Optogenetic Tuning of Protein-protein Binding in Bilayers Using LOVTRAP
    LOVTRAP对双层膜蛋白质结合的光遗传学调控
    作者:Doug Tischer and Orion D. Weiner日期:09/05/2020,浏览量:660,Q&A: 0
    [Abstract] Modern microscopy methods are powerful tools for studying live cell signaling and biochemical reactions, enabling us to observe when and where these reactions take place from the level of a cell down to single molecules. With microscopy, each cell or molecule can be observed both before and after a given perturbation, facilitating better inference ...
    Karyopherin-β2 Inhibits and Reverses Aggregation and Liquid-liquid Phase Separation of the ALS/FTD-Associated Protein FUS
    核转运蛋白-β2抑制和ALS / FTD相关蛋白FUS的逆转聚集和液-液相分离
    作者:Emma Robinson, James Shorter and Lin Guo日期:08/20/2020,浏览量:758,Q&A: 0
    [Abstract] The study of RNA-binding proteins (RBP) offers insight into the mechanisms of pathologic protein aggregation in neurodegenerative diseases. We developed a protocol for purifying an RBP FUS and a nuclear import receptor (NIR) Kapβ2 and testing the ability of Kapβ2 to mitigate FUS aggregation and liquid-liquid phase separation.
    Microtubule Seeded-assembly in the Presence of Poorly Nucleating Nucleotide Analogues
    存在不良成核核苷酸类似物的微管种晶组装
    作者:Siou Ku, Claire Heichette, Laurence Duchesne and Denis Chrétien日期:08/20/2020,浏览量:464,Q&A: 0
    [Abstract] Microtubule dynamic instability is driven by the hydrolysis of the GTP bound to the β-subunit of the α-β tubulin heterodimer. Nucleotide analogues are commonly used to mimic the different steps of the tubulin GTPase cycle, but most of them are poor microtubule nucleators. Usually, microtubule assembly is seeded by guanylyl-(α, ...
    Determination of the Cellular Ion Concentration in Saccharomyces cerevisiae Using ICP-AES
    ICP-AES法测定酿酒酵母细胞离子浓度
    作者:Louise Thines, Anne Iserentant and Pierre Morsomme日期:08/20/2020,浏览量:589,Q&A: 0
    [Abstract] The yeast Saccharomyces cerevisiae has been perceived over decades as a highly valuable model organism for the investigation of ion homeostasis. Indeed, many of the genes and biological systems that function in yeast ion homeostasis are conserved throughout unicellular eukaryotes to humans. In this context, measurement of the yeast ...
    The ATPase Activity of Escherichia coli Expressed AAA+-ATPase Protein
    大肠杆菌表达的AAA + -ATPase蛋白的ATPase活性
    作者:Amita Kaundal, Vemanna S. Ramu and Kirankumar S. Mysore日期:08/05/2020,浏览量:570,Q&A: 0
    [Abstract] ATPases are the enzymes that breakdown ATP to ADP and release inorganic phosphate (Pi). Here we provide a detailed protocol to determine the ATPase activity of a recombinant AAA+-ATPase protein (GENERAL CONTROL NON-REPRESSIBLE-4 [GCN4]) by spectrophotometric absorption at 360 nm to measure the accumulated inorganic phosphate. In ...
    A Method for User-defined Mutagenesis by Integrating Oligo Pool Synthesis Technology with Nicking Mutagenesis
    一种集成引物池合成技术和切口诱变的用户自定义突变方法
    作者:Paul J. Steiner, Zachary T Baumer and Timothy A. Whitehead日期:08/05/2020,浏览量:795,Q&A: 0
    [Abstract] Saturation mutagenesis is a fundamental enabling technology for protein engineering and epitope mapping. Nicking mutagenesis (NM) allows the user to rapidly construct libraries of all possible single mutations in a target protein sequence from plasmid DNA in a one-pot procedure. Briefly, one strand of the plasmid DNA is degraded using a nicking ...
    Expression and Purification of Functionally Active Serotonin 5-HT2A Receptor in Insect Cells Using Low-titer Viral Stock
    低滴度病毒载体在昆虫细胞中表达和纯化功能性5-HT2A受体
    作者:Sukanya Mozumder, Gopa Mahesh, Krishnamoorthi Srinivasan, Jayati Sengupta and Sujoy Mukherjee日期:08/05/2020,浏览量:665,Q&A: 0
    [Abstract] The serotonin 5-HT2A receptor (5-HT2AR) is a member of the GPCR family that is important for various neurological functions and whose dysregulation causes many mental health disorders. Structural investigations of 5-HT2AR require the production of functionally active receptors expressed from eukaryotic cell ...
    Production and Isolation of Magnetic Protein Crystals in HEK293T Cells
    HEK293T细胞中磁性蛋白晶体的生产与分离
    作者:Thomas L. Li and Bianxiao Cui日期:07/20/2020,浏览量:586,Q&A: 0
    [Abstract] Advances in protein engineering have enabled the production of self-assembled protein crystals within living cells. Our recent publication demonstrates the production of ftn-PAK4, which is a ferritin-containing crystal that can mineralize iron and become magnetic when isolated. We have developed an optimized protocol for the production and ...
    RNA Stability Measurements Using RT-qPCR in Arabidopsis Seedlings
    RT-qPCR法测定拟南芥幼苗RNA稳定性
    作者:Tianran Jia and Brandon H. Le日期:07/20/2020,浏览量:371,Q&A: 0
    [Abstract] Steady-state mRNA levels are determined by both the rates of transcription and degradation. Regulation of mRNA stability and/or degradation are key factors that can significantly affect mRNA levels and its biological functions. mRNA stability can be measured indirectly after transcription inhibition. This protocol described a rapid and sensitive ...