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Biotechnology and Bioprocess Engineering : Volume 5, Issue 1, February 2000

Biotechnology and Bioprocess Engineering : Volume 4

Biotechnology and Bioprocess Engineering : Volume 3

Biotechnology and Bioprocess Engineering : Volume 2

Permanent mycoplasma removal from tissue culture cells: A genetic approach

Content Provider	Springer Nature Link
Author	Mohr, Gabriele Preininger, Alexandra Himmelspach, Michèle Plaimauer, Barbara Arbesser, Christine York, Heinz Dorner, Friedrich Schlokat, Uwe
Copyright Year	2000
Abstract	Mycoplasma contamination of tissue culture cells easily evades detection and, thus, represents a continous threat to cell biologists. In cases where infected cell can not simply be replaced, attempts have to be made to eradicate mycoplasma from the tissue culture cells. A variety of anti-microbial agents have been shown to be toxic to mycoplasma strains; however, cell associated mycoplasmas are often protected from antibiotics at concentrations shown to be effectivein vitro. Antibiotic concentrations high enough to be lethal to cell asso\|ciated mycoplasmas frequently are also detrimental to the host cells, while moderately increased antibiotic levels tolerated by the host cells often lead to only temporary growth suppression and/or to the emergence of mycoplasma strains resistant even to high concentrations of the antibiotic applied. Here, a genetic approach for the elimination of mycoplasma from tissue culture cells that overcomes these limitations is described. By expression of a selection marker conferring resistance to an otherwise toxic agent,Acholeplasma laidlawii infected BHK-21 cells used as the model system were enabled to temporarily tolerate antibiotic concentrations high enough to be lethal to cell associated mycoplasma while leaving the host cells unharmed. Upon successful mycoplasma eradication, cultivation of the cured host cells in the absence of the selective agent yielded revertant cell clones that had regained susceptibility to the toxic agent. Cessation of the selection marker expression was shown to result from the loss of the selection marker DNA, which is a consequence of the fact that the stable and permanent integration of foreign DNA in eucaryotic cell chromosomes is highly inefficient. Thus, the cells were cured from mycoplasma yet remained biochemically unaltered.
Starting Page	84
Ending Page	91
Page Count	8
File Format	PDF
ISSN	12268372
Journal	Biotechnology and Bioprocess Engineering
Volume Number	5
Issue Number	2
e-ISSN	19763816
Language	English
Publisher	The Korean Society for Biotechnology and Bioengineering
Publisher Date	2000-01-01
Publisher Place	Seoul
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	mycoplasma selection cell culture techniques Biotechnology Industrial and Production Engineering
Content Type	Text
Resource Type	Article
Subject	Bioengineering Biomedical Engineering Applied Microbiology and Biotechnology Biotechnology

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