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Controversial aspect of using GFP as a marker for the production of transgenic cattle

100%

Duszewska A. M., Lipinski D., Piliszek A., Slomski R., Plawski A., Wojdan J., Gawron W., Juzwa W., Zeyland J., Wenta-Muchalska E., Reklewski Z.

Polish Journal of Veterinary Sciences

2004

tom 07

nr 4

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 (hTNFa) gene fused to the bovine betalactoglobulin promoter (bLG) in plasm id vector pCX-EGFP. In four experiments, 76 zygotes were injected; eight of them developed to the morulac/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.

Ograniczanie wyników

1 Polish Journal of Veterinary Sciences

1 Duszewska A. M.

1 Gawron W.

1 Juzwa W.

1 Lipinski D.

1 Piliszek A.

1 Plawski A.

1 Reklewski Z.

1 Slomski R.

1 Wenta-Muchalska E.

1 Wojdan J.

1 Zeyland J.

1 2004

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Controversial aspect of using GFP as a marker for the production of transgenic cattle