ORIGINAL PAPER
The use of green fluorescent protein (GFP)
to select bovine embryos
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1
Institute of Genetics and Animal Breeding, Polish Academy of Sciences,
Experimental Embryology Department, Jastrzębiec, 05-552 Wólka Kosowska, Poland
2
All Russian Research Institute for Farm Animal Genetics and Breeding,
55a Moskovskoe shosse, St. Petersburg-Pushkin, 189620 Russia
3
Institute of Genetics and Animal Breeding, Polish Academy of Sciences,
Molecular Cytogenetics Department, Jastrzębiec, 05-552 Wólka Kosowska, Poland
4
Białystok Technical University, Faculty of Building and Environmental Engineering,
Chair of Sanitary Biology and Biotechnology,
Wiejska 45a, 15-351 Białystok, Poland
Publication date: 2003-01-02
J. Anim. Feed Sci. 2003;12(1):71-81
KEYWORDS
ABSTRACT
A major factor limiting the transgenesis in domestic animals is the inefficiency of maintaining
large numbers of recipients carrying nontransgenic foetuses. The objectives of this study were: 1. to
determine the influence of green fluorescent protein (GFP) construct injection on the development of
bovine embryos, 2. to identify and select the GFP positive bovine embryos, and 3. to determine the
rate of mosaicism in transgenic embryos. Cattle oocytes were matured and fertilised in vitro and zygotes were microinjected with pCX-EGFP construct consisting of CMV-IE enhancer, chicken β-actin promoter, cDNA of GFP (EGFP-732 bp) and rabbit β-globin polyadenylation sequences. Embryos
from control (64) and microinjected (198) groups were cultured in vitro. After 168 h of culture, morula and blastocysts were observed in 39.06% of control and in 23.23% of injected group. We obtained three GFP positive embryos (1.51% of injected zygotes and 6.52% of morulae/blastocysts). One
of them was 100, second 75 and third 25% GFP positive (66.7% of mosaicism). Use of gfp gene reporter to select bovine embryos is useful method to increase transgenic offspring, because GFP marker allows to choice only transgenic embryos and transfer them to recipients.
CITATIONS (1):
1.
Pre-Implantation Bovine Embryo Evaluation—From Optics to Omics and Beyond
R. Rabel, Paula Marchioretto, Elizabeth Bangert, Kenneth Wilson, Derek Milner, Matthew Wheeler
Animals