title: Controversial aspect of using GFP as a marker for the production of transgenic cattle.
abstract: The objective of this study was to examine the feasibility of identification and selection of cattle embryos based on green fluorescence (GFP-positive) in order to obtain calves carrying an integrated transgene. The construct used (pbLGTNF-EGFP) contained the human tumor necrosis factor alpha (hTNFalpha) gene fused to the bovine beta-lactoglobulin promoter (bLG) in plasmid vector pCX-EGFP. In four experiments, 76 zygotes were injected; eight of them developed to the morulae/blastocysts stage of which only five were GFP positive (one of them 100%, one-50%, three- 25%). All of the GFP positive embryos were transferred to recipients. Two calves were born: one after transfer of the 100% GFP positive embryo and the other after transfer of one of the 25% GFP positive embryos. Both animals were healthy with normal weight when compared to two control calves. The integration of pbLGTNF-EGFP in the host genome could not be detected in either of the calves, suggesting that GFP is an unreliable marker for preimplantation screening of embryos.
authors.affiliation: Department of Experimental Embryology, Polish Academy of Sciences, Strzeszyńska 32, 60-479 Poznań, Poland. duszewskaanna@hotmail.com
authors.forename: A M
authors.initials: AM
authors.lastname: Duszewska
authors.forename: D
authors.initials: D
authors.lastname: Lipiński
authors.forename: A
authors.initials: A
authors.lastname: Piliszek
authors.forename: R
authors.initials: R
authors.lastname: Słomski
authors.forename: A
authors.initials: A
authors.lastname: Pławski
authors.forename: J
authors.initials: J
authors.lastname: Wojdan
authors.forename: W
authors.initials: W
authors.lastname: Gawron
authors.forename: W
authors.initials: W
authors.lastname: Juzwa
authors.forename: J
authors.initials: J
authors.lastname: Zeyland
authors.forename: E
authors.initials: E
authors.lastname: Wenta-Muchalska
authors.forename: Z
authors.initials: Z
authors.lastname: Reklewski
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D015415:Biomarkers; D001755:Blastocyst; D002417:Cattle / Q000235:genetics*; D004624:Embryo Transfer / Q000662:veterinary; D004622:Embryo, Mammalian / Q000166:cytology* / Q000378:metabolism; D005240:Feasibility Studies; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D007782:Lactoglobulins / Q000235:genetics; D008845:Microinjections / Q000662:veterinary; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011993:Recombinant Fusion Proteins / Q000235:genetics; D019076:Transgenes / Q000235:genetics; D014180:Transplantation / Q000662:veterinary; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
chemical_list: D015415:Biomarkers; D007782:Lactoglobulins; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins