Publications associées à Plasmide Ti

Two defence systems eliminate plasmids from seventh pandemic Vibrio cholerae

Melanie Blokesch, David William Adams, Milena Jadwiga Jaskolska

Horizontal gene transfer can trigger rapid shifts in bacterial evolution. Driven by a variety of mobile genetic elements—in particular bacteriophages and plasmids—the ability to share genes within and across species underpins the exceptional adaptability o ...

2022

Engineering synthetic adhesins for biophysical investigation and rewiring of host-microbe interactions

Xavier Jean-Yves Pierrat

Host attachment is often a critical step in the onset of pathogenesis. To attach to host cells, bacteria have evolved a range of adhesins that bind to specific receptors. Some of these adhesins have been thoroughly characterized using biochemical technique ...

EPFL2022

The studies of ParA and ParB dynamics reveal asymmetry of chromosome segregation in mycobacteria

John McKinney, Isabella Santi, Djenet Bousbaine

Active segregation of bacterial chromosomes usually involves the action of ParB proteins, which bind in proximity of chromosomal origin (oriC) regions forming nucleoprotein complexes - segrosomes. Newly duplicated segrosomes are moved either uni- or bidire ...

Wiley2017

Study of transposon-mediated cell pool and cell line generation in CHO cells

Sowmya Balasubramanian

The goal of this thesis was to evaluate the use of artificial transposon systems for the generation of recombinant cell pools and cell lines with Chinese hamster ovary (CHO) cells as the production host. Transposons are naturally occurring genetic elements ...

EPFL2015

Effects of physiological self-crowding of DNA on shape and biological properties of DNA molecules with various levels of supercoiling

Giovanni Dietler, Julien Dorier, Fabrizio Benedetti, Yannis Burnier, Aleksandre Japaridze, Andrzej Stasiak, Robert Sebastian Kwapich

DNA in bacterial chromosomes and bacterial plasmids is supercoiled. DNA supercoiling is essential for DNA replication and gene regulation. However, the density of supercoiling in vivo is circa twice smaller than in deproteinized DNA molecules isolated from ...

Oxford University Press2015

Development of plasmid-based expression technology for rapid and sustained production of recombinant proteins in insect cells

Xiao Shen

Due to the high demand for heterogeneous protein production in insect cells, we have established efficient methods for plasmid-based transient gene expression (TGE) in various insect cell lines, including commonly used Spodoptera cell lines Sf9 and HighFiv ...

EPFL2014

MAR-mediated integration of plasmid vectors for in vivo gene transfer and regulation

Andrzej Kulik, Nicolas Mermod, Stefania Puttini, Déborah Ley, Damien Saugy

Background: The in vivo transfer of naked plasmid DNA into organs such as muscles is commonly used to assess the expression of prophylactic or therapeutic genes in animal disease models. Results: In this study, we devised vectors allowing a tight regulatio ...

BioMed Central2013

TransFLP - A Method to Genetically Modify Vibrio cholerae Based on Natural Transformation and FLP-recombination

Melanie Blokesch

Several methods are available to manipulate bacterial chromosomes(1-3). Most of these protocols rely on the insertion of conditionally replicative plasmids (e.g. harboring pir-dependent or temperature-sensitive replicons(1,2)). These plasmids are integrate ...

2012

Calcium phosphate transfection generates mammalian recombinant cell lines with higher specific productivity than polyfection

Florian Maria Wurm, David Hacker, Madiha Derouazi, Sébastien Chenuet, Nicolas Jaccard

Transfection with polyethylenimine (PEI) was evaluated as a method for the generation of recombinant Chinese hamster ovary (CHO DG44) cell lines by direct comparison with calcium phosphate-DNA coprecipitation (CaPO4) using both green fluorescent protein (G ...

2008

Accelerating the development time of recombinant mammalian cell lines producing high titers of protein therapeutics

Sébastien Chenuet

The forthcoming arrival on the market of numerous protein therapeutics that require high clinical doses will foster the need for high-producing mammalian cell lines. Furthermore, pressures to reduce the development time for new therapeutics has meant more ...

EPFL2008

Multiple glycerol shocks increase the calcium phosphate transfection of non-synchronized CHO cells

Florian Maria Wurm, David Hacker, Martin Bertschinger, Frédéric Grosjean

The exposure of CHO DG44 cells to an osmotic shock, after DNA uptake, results in a cellular volume decrease of approx. 55%. Repetitive osmotic shocks targeted different sub-populations of cells as was demonstrated using two different fluorescent reporter g ...

2006

The use of RNA interference to increase PEI-mediated transient gene expression in mammalian cells

Martin Bertschinger

DNA uptake by polyethylenimine (PEI)-mediated transfection was investigated in Chinese hamster ovary (CHO DG44) cells. Rapid DNA uptake was observed with the maximum occurring during the first 45-60 min after addition of PEI-DNA complexes to cells. With lo ...

EPFL2006

Genetic characterization of CHO production host DG44 and derivative recombinant cell lines

Florian Maria Wurm, David Hacker, Madiha Derouazi, Martin Bertschinger

The dihydrofolate reductase-deficient Chinese hamster ovary (CHO) cell line DG44 is the dominant mammalian host for recombinant protein manufacturing, in large part because of the availability of a well-characterized genetic selection and amplification sys ...

2006

Establishment of recombinant mammalian cell lines : new approaches

Madiha Derouazi

Recombinant proteins are gaining in importance for therapeutic applications. The proteins are expressed in stable cell lines, within which recombinant DNA has incorporated into the host cells genome. Identification and isolation of extremely high producers ...

EPFL2005