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Green fluorescent protein (GFP) transsenic pig produced by somatic cell nuclear transfer
作者姓名: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
作者单位:[1]Life Science College, North-east Agricultural University, Harbin 150030, China; [2]Animal Medical College, Northeast Agricultural University, Harbin 150030, China; [3]Harbin Sanyuan Animal Husbandry Co. Ltd., Harbin 150030, China; [4]Division of Animal Science, University of Missouri-Columbia, Columbia MO 65211, USA
摘    要: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 transfecUon 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 resulted in the highest transfecUon 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%. 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.

关 键 词:GFP  体细胞  核子传输  遗传学
收稿时间:2007-03-29

Green fluorescent protein (GFP) transgenic pig produced by somatic cell nuclear transfer
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.Green fluorescent protein (GFP) transgenic pig produced by somatic cell nuclear transfer[J].Chinese Science Bulletin,2008,53(7):1035-1039.
Authors:ZhongHua Liu  Jun Song  ZhenKun Wang  JiangTian Tian  QingRan Kong  Zhong Zheng  Zhi Yin  Li Gao  HaiKun Ma  Shuang Sun  YuTian Li  HongBin Wang  R S Prather
Institution:(1) Life Science College, North-east Agricultural University, Harbin, 150030, China;(2) Animal Medical College, Northeast Agricultural University, Harbin, 150030, China;(3) Harbin Sanyuan Animal Husbandry Co. Ltd., Harbin, 150030, China;(4) Division of Animal Science, University of Missouri-Columbia, Columbia, MO 65211, USA
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. Contributed equally to this work
Keywords:transgenic  green fluorescent protein  nuclear transfer  liposome  pig
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