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

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  • Development Biology
  • Published:
Chinese Science Bulletin

Abstract

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 development of reconstructed embryos both in vitro and in vivo was 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 h 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 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.

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Authors and Affiliations

  1. Life Science College, North-east Agricultural University, Harbin, 150030, China

    ZhongHua Liu, Jun Song, ZhenKun Wang, JiangTian Tian, QingRan Kong, Zhong Zheng, Zhi Yin & Shuang Sun

  2. Animal Medical College, Northeast Agricultural University, Harbin, 150030, China

    Li Gao, HaiKun Ma & HongBin Wang

  3. Harbin Sanyuan Animal Husbandry Co. Ltd., Harbin, 150030, China

    YuTian Li

  4. Division of Animal Science, University of Missouri-Columbia, Columbia, MO, 65211, USA

    R. S. Prather

Authors
  1. ZhongHua Liu
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  2. Jun Song
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  3. ZhenKun Wang
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  4. JiangTian Tian
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  5. QingRan Kong
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  6. Zhong Zheng
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  7. Zhi Yin
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  8. Li Gao
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  9. HaiKun Ma
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  10. Shuang Sun
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  11. YuTian Li
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  12. HongBin Wang
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  13. R. S. Prather
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Corresponding author

Correspondence to ZhongHua Liu.

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Contributed equally to this work

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Liu, Z., Song, J., Wang, Z. et al. Green fluorescent protein (GFP) transgenic pig produced by somatic cell nuclear transfer. Chin. Sci. Bull. 53, 1035–1039 (2008). https://doi.org/10.1007/s11434-008-0168-x

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  • DOI: https://doi.org/10.1007/s11434-008-0168-x

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