Publication

The Semliki Forest virus system: receptor expression and scale-up

Horst Arthur Vogel, Rudolf Hovius
1997
Journal Articles

Abstract

The highly efficient Semliki Forest virus (SFV) system has been applied to express several recombinant 7TM receptors and ligand-gated ion channels. High affinity binding and functional coupling to G proteins were obtained for the receptors. The ligand-gated ion channels showed high specific binding activity and high-amplitude electrophysiol. responses in whole-cell patch clamp recordings. The rapid and simple generation of high titer virus stocks has made the SFV system also useful for site-directed mutagenesis studies. The broad host range of SFV has allowed studies of recombinant proteins in a wide variety of cell lines and primary cell cultures. Scale-up technol. for SFV has been established. Recombinant SFV particles contg. a his-tagged mouse serotonin 5-HT3 receptor gene were used to infect suspension cultures of BHK cells at reactor scale (11.5 L). Receptor yields of 15 mg (Bmax values of 52 pmol mg-1) were obtained. The receptor was solubilized, isolated to high purity and subjected to structure-function studies. [on SciFinder (R)]

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https://infoscience.epfl.ch/entities/publication/66f1babc-c43d-4543-bd87-9bbd19e1743e

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Horst Arthur Vogel, Rudolf Hovius
1997
Journal Articles

Abstract

Official source

https://infoscience.epfl.ch/entities/publication/66f1babc-c43d-4543-bd87-9bbd19e1743e

About this result

Related concepts (32)

Cell culture

Cell culture or tissue culture is the process by which cells are grown under controlled conditions, generally outside of their natural environment. The term "tissue culture" was coined by American pathologist Montrose Thomas Burrows. This technique is also called micropropagation. After the cells of interest have been isolated from living tissue, they can subsequently be maintained under carefully controlled conditions. They need to be kept at body temperature (37 °C) in an incubator.

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A stem cell line is a group of stem cells that is cultured in vitro and can be propagated indefinitely. Stem cell lines are derived from either animal or human tissues and come from one of three sources: embryonic stem cells, adult stem cells, or induced stem cells. They are commonly used in research and regenerative medicine. Stem cell By definition, stem cells possess two properties: (1) they can self-renew, which means that they can divide indefinitely while remaining in an undifferentiated state; and (2) they are pluripotent or multipotent, which means that they can differentiate to form specialized cell types.

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Primary cell culture is the ex vivo culture of cells freshly obtained from a multicellular organism, as opposed to the culture of immortalized cell lines. In general, primary cell cultures are considered more representative of in vivo tissues than cell lines, and this is recognized legally in some countries such as the UK (Human Tissue Act 2004). However, primary cells require adequate substrate and nutrient conditions to thrive and after a certain number of divisions they acquire a senescent phenotype, leading to irreversible cell cycle arrest.

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The Semliki Forest virus system: receptor expression and scale-up

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