Transient Expression

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Alain Bernard - One of the best experts on this subject based on the ideXlab platform.

  • large scale Transient Expression in mammalian cells for recombinant protein production
    Current Opinion in Biotechnology, 1999
    Co-Authors: Florian M Wurm, Alain Bernard

    Large-scale Transient Expression from mammalian cells is a new technology. Breakthroughs have been achieved for non-viral delivery methods: transfections can now be done at the 1-10 L scale with mammalian cells grown in suspension. Production of 1-20 mg/L of recombinant protein have been obtained in stirred bioreactors. Modified alphaviruses have provided a fast and efficient Expression technology based on viral vectors.

  • Large scale Transient Expression with COS cells.
    Cytotechnology, 1996
    Co-Authors: Horst Blasey, Jean-pierre Aubry, Gonzalo J. Mazzei, Alain Bernard

    We demonstrate here that Transient Expression with COS cells can be performed at the one litre scale for a period of more than 10 days. Cells grown in T225 flasks were transfected by electroporation, transferred into spinners, and then grown either in suspension or on microcarriers. A daily medium change significantly extented culture life and production time, compared with standard protocols.

Florian M Wurm - One of the best experts on this subject based on the ideXlab platform.

Guoliang Wang - One of the best experts on this subject based on the ideXlab platform.

  • rice oryza sativa protoplast isolation and its application for Transient Expression analysis
    Current protocols in plant biology, 2016
    Co-Authors: Feng He, Songbiao Chen, Yuese Ning, Guoliang Wang

    Rice (Oryza sativa) is not only the staple food for half of the world's population but also a model monocot plant for molecular biology studies. Although rice genes have been extensively investigated in the last two decades, the functions of many genes in the rice genome are still not known. One of the rapid and efficient approaches for determining gene function in vivo is protoplast-based Transient Expression analysis. We established a rice protoplast system about 10 years ago, which has been recently used in many laboratories. This protocol is useful for protein Expression, subcellular localization, bimolecular fluorescence complementation, and co-immunoprecipitation assays. © 2016 by John Wiley & Sons, Inc. Keywords: rice (Oryza sativa); protoplast isolation; Transient Expression; BiFC and Co-IP

Craig S. Pikaard - One of the best experts on this subject based on the ideXlab platform.

  • Transient Expression in Arabidopsis thaliana protoplasts derived from rapidly established cell suspension cultures
    Plant Cell Reports, 1993
    Co-Authors: Jed H. Doelling, Craig S. Pikaard

    Arabidopsis thaliana has emerged as a model species for the analysis of genes controlling plant development. However, its small size has impaired biochemical analyses, and the absence of a Transient Expression system has hampered promoter analysis. Here, we report a method for rapidly establishing A. thaliana suspension cultures that yield protoplasts that can be readily transfected. We have optimized Transient Expression conditions using a modified polyethylene glycol / calcium nitrate transformation protocol and a Cauliflower Mosaic Virus 35S promoter-luciferase reporter gene construct. Our methods permit isolation of large quantities of rapidly growing cells and analysis of Arabidopsis promoters in vivo in a homologous system.

Noboru Murase - One of the best experts on this subject based on the ideXlab platform.

  • Reporter gene introduction and Transient Expression in protoplasts of Porphyra yezoensis
    Journal of Applied Phycology, 2004
    Co-Authors: Yuzuru Mizukami, Makoto Hado, Hitoshi Kito, Masahiko Kunimoto, Noboru Murase

    The Transient Expression of foreign genes in the protoplasts of Porphyrayezoensis was examined using three recombinant vectors, pYez-Rub-GUS, pYez-Rub-GFP and pYez-Rub-LUC, which were constructed with the promoter sequence of the ribulose-bisphosphate-carboxylase / oxygenase (Rubisco) gene as a promoter and the bacterial β-glucuronidase (GUS), mutant of green fluorescent protein (S65T-GFP) and firefly luciferase (LUC) genes, respectively, as reporter genes. When the pYez-Rub-GUS was introduced into protoplasts by electroporation, cells stained dark blue by indigotin were observed after the histochemical GUS assay. GUS activity was also detected by quantitative enzyme assays with a chemiluminescent substrate. When the pYez-Rub-GFP was electroporated into protoplasts, the Expression of GFP could be detected in vivo observations with fluorescence microscopy. However, the rates of gene Expression cells to the total number of cells were different between the GUS and GFP genes. LUC activity was also detected by assay with a chemiluminescent substrate after the introduction of pYez-Rub-LUC into protoplasts, although the activity levels were considerably lower. Relatively high Expression rates of introduced GUS genes were observed 3 to 5 days after electroporation. These results show that the promoter sequence of the chloroplast Rubisco gene functions as a promoter of foreign gene Expression and that Transient Expression occurred in protoplasts of P. yezoensis after the introduction of foreign genes.