Application of Green Fluorescent Protein as a Marker for Selection of Transgenic Mouse Embryos before Implantation

Authors : Haydar BAĞIŞ

Pages : 123-131

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Publication Date : 2001-12-01

Article Type : Research Paper

Abstract :In this study, we evaluated the application of green fluorescent protein insert ignore into journalissuearticles values(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 insert ignore into journalissuearticles values(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 insert ignore into journalissuearticles values(FITC); filter set attached to a microscope. In this analysis, 34 % of the two-cell stage embryos insert ignore into journalissuearticles values(37 embryos out of 108); were found to be GFP positive. Of these GFP-positive embryos, 8 embryos out of 37 insert ignore into journalissuearticles values(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