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KMID : 1005220120270010009
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Journal of Embryo Transfer
2012 Volume.27 No. 1 p.9 ~ p.14
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Production of α1,3-Galactosyltransferase (GalT) Double Knock-out (-/-) Transgenic Pigs for Xenotransplantation
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Hwnag Seong-Soo
Oh Keon-Bong
Kim Dong-Hoon
Woo Jae-Seok
Shim Ho-Sup
Yun Ik-Jin
Park Jin-Ki
Im Gi-Sun
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Abstract
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This study was conducted to analyze the transgenic efficiency and sex ratio in α-1,3-galactosyltransferase (GalT) knock-out (KO) transgenic pigs according to generation. GalT KO piglets were produced by artificial insemination or natural mating. The transgenic confirmation of GalT KO was evaluated by PCR amplification using specific primers. After electrophoresis, three types of bands were detected such as 2.3 kb single band (Wild), 2.3 and 3.6kb double bands (GalT KO ?/+; heterozygote), and 3.6kb single band (GalT KO ?/?; homozygote). Transgenic efficiency in F1 generation was 64.5% (23/35) of GalT KO (?/+). In F2 generation, GalT KO transgenic efficiency was 36.4% (21/57, Wild), 47.5% (28/57, GalT KO ?/+), and 16.1% (8/57, GalT KO ?/?), respectively. Interestingly, no ho- mozygote piglets were born in 6 deliveries among total 11 deliveries, although they were pregnant between male (M) and female (F) F1 heterozygote. In the 5 litters including at least one GalT KO ?/? piglet, the transgenic efficiency was 13.3% (2/24, Wild), 51.3% (14/24, GalT KO ?/+), and 35.3% (8/24, GalT KO ?/?), respectively. The sex ratio of M and F was 40:60 in F1 and 49:51 in F2 generation, respectively. Based on these results, GalT KO transgenic pigs have had a reproductive ability with a normal range of transgenic efficiency and sex ratio.
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KEYWORD
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α-1,3-galactosyltransferase , gene knock-out , artificial insemination , transgenic cloned pig , transgenic efficiency
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