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

Chinese Science Bulletin 2008年第7期53卷 1035-1039页

作者： LIU ZhongHua SONG Jun WANG ZhenKun TIAN JiangTian KONG QingRan ZHENG Zhong YIN Zhi GAO Li MA HaiKun SUN Shuang LI YuTian WANG HongBin R S PRATHER Life Science College North-east Agricultural University Harbin 150030 China Animal Medical College Northeast Agricultural University Harbin 150030 China Harbin Sanyuan Animal Husbandry Co. Ltd. Harbin 150030 China Division of Animal Science University of Missouri-Columbia Columbia MO 65211 USA

Transgenic somatic cell nuclear transfer is a very promising route for producing transgenic farm ani-mals. 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 development of reconstructed embryos both in vitro and in vivo was observed, and GFP expression was determined. The results showed that porcine fe-tal-derived fibroblast cells cultured with 4.0 μL/mL liposome and 1.6 μg/mL plasmid DNA for 6 h re-sulted in the highest transfection rate (3.6%). The percentage of GFP reconstructed embryos that de-veloped in vitro to the blastocyst stage was 10%. Of those the GFP positive percentage was 48%. Re-constructed transgenic embryos were transferred to 10 recipients. 5 of them were pregnant, and 3 de-livered 6 cloned piglets 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.

关键词： GFP 体细胞核子传输遗传学

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