Expression of the green fluorescent protein in Paramecium tetraurelia

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2000
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Hauser, Karin
Haynes, W. John
Kung, Ching
Plattner, Helmut
Kissmehl, Roland
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European Journal of Cell Biology. 2000, 79(2), pp. 144-149. ISSN 0171-9335. Available under: doi: 10.1078/S0171-9335(04)70016-3
Zusammenfassung

In this paper we describe the expression of green fluoresent protein (GFP) as a reporter in vivo to monitor transformation in Paramecium cells. This is not trivial because of the limited number of strong promoters available for heterologous expression and the very high AT content of the genomic DNA, the consequence of which is a very aberrant codon usage. Taking into account differences in codon usage we selected and modified the original GFP open reading frame (ORF) from Aequorea victoria and placed the altered ORF into the Paramecium expression vector pPXV. Injection of the linearized plasmid into the macronucleus resulted in a cytoplasmic fluorescence signal in the clonal descendants, which was proportional to the number of copies injected. Southern hybridization indicated the establishment and replication of the plasmid during vegetative growth. Expression was also monitored by Northern and Western analysis. The results indicate that the modified GFP can be used in Paramecium as a reporter for transformation as an alternative to selection with antibiotics and that it may also be used to construct and localize fusion proteins.

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570 Biowissenschaften, Biologie
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GFP, expression, Paramecium, transformation
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ISO 690HAUSER, Karin, W. John HAYNES, Ching KUNG, Helmut PLATTNER, Roland KISSMEHL, 2000. Expression of the green fluorescent protein in Paramecium tetraurelia. In: European Journal of Cell Biology. 2000, 79(2), pp. 144-149. ISSN 0171-9335. Available under: doi: 10.1078/S0171-9335(04)70016-3
BibTex
@article{Hauser2000Expre-6612,
  year={2000},
  doi={10.1078/S0171-9335(04)70016-3},
  title={Expression of the green fluorescent protein in Paramecium tetraurelia},
  number={2},
  volume={79},
  issn={0171-9335},
  journal={European Journal of Cell Biology},
  pages={144--149},
  author={Hauser, Karin and Haynes, W. John and Kung, Ching and Plattner, Helmut and Kissmehl, Roland}
}
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