- Turkish Journal of Biology
- Vol: 25 Issue: 2
- Application of Green Fluorescent Protein as a Marker for Selection of Transgenic Mouse Embryos befor...
Application of Green Fluorescent Protein as a Marker for Selection of Transgenic Mouse Embryos before Implantation
Authors : Haydar Bağiş
Pages : 123-131
View : 9 | Download : 5
Publication Date : 9999-12-31
Article Type : Makaleler
Abstract :In this study, we evaluated the application of green fluorescent protein (GFP) from the jellyfish Aequorea victoria, for efficient selection of possible transgenic and non-mosaic mouse embryos after microinjection. We injected 353 one-cell mouse zygotes with a DNA fragment carrying Gfp gene under the control of b-actin gene promoter. Eighty-seven per cent (307 embryos) of the injected embryos survived after microinjection. The surviving embryos were cultured for an additional day and then analysed for detection of GFP expression using a standard fluorescein isothiocyanate (FITC) filter set attached to a microscope. In this analysis, 34 % of the two-cell stage embryos (37 embryos out of 108) were found to be GFP positive. Of these GFP-positive embryos, 8 embryos out of 37 (21%) were found to be non-mosaic as GFP expression was detected in all the blastomeres. Our results show that the successful use of GFP as a vital marker for detection of mosaic embryos holds promise for potential application of this technology as a selection tool in the generation of transgenic animals.Keywords : Transgenic mice, embryo, microinjection, green fluorescent protein