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    Protocols in Current Issue
    Visualization and Quantification of Cell-to-cell Movement of Proteins in Nicotiana benthamiana
    Authors:  Mila C Blekemolen, Mariliis Tark-Dame and Frank L.W Takken, date: 12/20/2018, view: 1185, Q&A: 0
    [Abstract] Cell-to-cell movement of proteins through plasmodesmata is a widely-established mechanism for intercellular signaling in plants. Current techniques to study intercellular protein translocation rely on single-cell transformation using particle bombardment or transgenic lines expressing photo-inducible fluorophores. The method presented here allows ...
    Ubiquitin Proteasome Activity Measurement in Total Plant Extracts
    Authors:  Suayib Üstün and Frederik Börnke, date: 09/05/2017, view: 3737, Q&A: 0
    [Abstract] The fine-tuned balance of protein level, conformation and location within the cell is vital for the dynamic changes required for a cell to respond to a given stimulus. This requires the regulated turnover of damaged or short-lived proteins through the ubiquitin proteasome system (UPS). Thus, the protease activity of the proteasome is adjusted to ...
    Expression and Partial Purification of His-tagged Proteins in a Plant System
    Author:  Amancio de Souza, date: 09/05/2015, view: 6966, Q&A: 0
    [Abstract] Plant protein expression can be a challenging enterprise in any biochemical or molecular biology research project. Several heterologous systems like bacteria, yeast, insect cells and cell free systems have been used to produce plant proteins for in vitro experiments and structural characterization. However, due to particularities of plant ...
    Protein Immunoprecipitation Using Nicotiana benthamiana Transient Expression System
    Authors:  Fang Xu, Charles Copeland and Xin Li, date: 07/05/2015, view: 11786, Q&A: 0
    [Abstract] Nicotiana benthamiana (N. benthamiana) is a useful model system to transiently express protein at high level. This protocol describes in detail how to transiently express protein in N. benthamiana and how to carry out protein immunoprecipitation in this expression system. This protocol can be broadly used for ...
    Localization and Topology of Thylakoid Membrane Proteins in Land Plants
    Authors:  Salar Torabi, Magdalena Plöchinger and Jörg Meurer, date: 12/20/2014, view: 7385, Q&A: 0
    [Abstract] Thylakoids are a formation of flattened membrane vesicles and protein complexes found in cyanobacteria, algae and plants. In the chloroplasts of land plants the thylakoid membrane systems form a network of densely packed stacks called grana lamellae, which are connected by unstacked stroma lamellae. Photosystem II is mainly localized in the ...
    Fluorescence Recovery after Photobleaching (FRAP) Assay to Measure the Dynamics of Fluorescence Tagged Proteins in Endoplasmic Reticulum Membranes of Plant Cells
    Authors:  José Antonio Navarro, Marta Serra-Soriano and Vicente Pallás, date: 10/20/2014, view: 9746, Q&A: 0
    [Abstract] In this protocol, we used fluorescence recovery after photobleaching (FRAP) to measure the influence that some mutations and drug treatment have on mobility of a green fluorescent protein (GFP)-fused viral transmembrane protein into endoplasmic reticulum membranes (Serra-Soriano et al., 2014). The proteins of interest were transiently ...
    Extraction of Chloroplast Proteins from Transiently Transformed Nicotiana benthamiana Leaves
    Author:  Joern Klinkenberg, date: 09/20/2014, view: 11476, Q&A: 0
    [Abstract] This rapid protocol allows the extraction of chloroplast enriched proteins from Nicotiana benthamiana (N. benthamiana) leaves that were transiently transformed to express an epitope tagged protein of interest. Thus, it can serve as a tool to study the chloroplastidic localization of the protein of interest when it is combined ...
    SGR-based Reporter to Assay Plant Transcription Factor-promoter Interactions
    Authors:  Shisong Ma and Savithramma Dinesh-Kumar, date: 08/20/2014, view: 6519, Q&A: 0
    [Abstract] We developed an in vivo method to assay plant transcription factor (TF)–promoter interactions using the transient expression system in Nicotiana benthamiana (N. benthamiana) plants. The system uses the Arabidopsis stay green (SGR) gene as a reporter. Induction of SGR expression in N. ...
    Monoclonal Antibody Purification (Nicotiana benthamiana Plants)
    Authors:  Adam Husk, Krystal Teasley Hamorsky and Nobuyuki Matoba, date: 01/20/2014, view: 9036, Q&A: 1
    [Abstract] Plant-based expression systems provide an alternative biomanufacturing platform for recombinant proteins (Matoba et al., 2011). In particular, plant virus-based vectors can overexpress proteins within days in the leaf tissue of Nicotiana benthamiana (N. benthamiana). To overcome the issues of genetic instability and ...
    Subcellular Localization Experiments and FRET-FLIM Measurements in Plants
    Authors:  Malgorzata Lichocka and Elmon Schmelzer, date: 01/05/2014, view: 10258, Q&A: 0
    [Abstract] Determining the localization of proteins within living cells may be very essential for understanding their biological function. Usually for analysis of subcellular localization, a construct encoding the translational fusion of a cDNA of interest with a fluorescent protein (FP) is engineered, transiently expressed in plant cells and examined with ...



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