第４回　生命資源研究･支援センター国際シンポジウム

「Green fluorescent protein (GFP) transgenic pig produced by somatic cell nuclear transfer」

東北農業大学　生命科学技術学院　教授
熊本大学　生命資源研究・支援センター　客員教授　劉　忠華

　Transgenic somatic cell nuclear transfer is a very promising route for producing transgenic farm animals. Research on GFP transgenic pigs can provide useful information for breeding transgenic pigs, human disease models and human organ xenotransplantation. In this study, a liposomal transfection system was screened and transgenic embryos were reconstructed by nuclear transfer of GFP positive cells into enucleated in vitro matured oocytes. The developments of reconstructed embryos both in vitro and in vivo were observed, and GFP expression was determined. The results showed that porcine fetal-derived fibroblast cells cultured with 4.0 μl/ml liposome and 1.6 μg/ml plasmid DNA for 6 hours resulted in the highest transfection rate (3.6%). The percentage of GFP reconstructed embryos that developed in vitro to the blastocyst stage was 10%. Of those the GFP positive percentage was 48%. Reconstructed transgenic embryos were transferred to 10 recipients. 5 of them were pregnant, and 3 delivered 6 cloned p iglets in which 4 piglets were transgenic for the GFP as verified by both GFP protein expression and GFP DNA sequence analysis. The percentage of reconstructed embryos that resulted in cloned piglets was 1.0%; while the percentage of piglets that were transgenic was 0.7%. This is the first group of transgenic cloned pigs born in China, marking a great progress in Chinese transgenic cloned pig research.