title: Cloning of the transgenic pigs expressing human decay accelerating factor and N-acetylglucosaminyltransferase III.
abstract: The present paper describes production of cloned pigs from fibroblast cells of transgenic pigs expressing human decay accelerating factor (DAF, CD55) and N-acetylglucosaminyltransferase III (GnT-III) that remodels sugar-chain biosynthesis. Two nuclear transfer protocols were used: a two-step activation (TA) method and a delayed activation (DA) method. Enucleated in vitro-matured oocytes and donor cells were electrically fused in a calcium-containing medium by TA method or in a calcium-free medium by DA method, followed by electrical activation 1-1.5 h later, respectively. In vitro blastocyst formation rates of nuclear transferred embryos reconstructed by TA and DA method were 8% and 14%, respectively. As a result of embryo transfer of the reconstructed embryos made by each method into recipient pigs, both gave rise to cloned piglets. These cloned pigs expressed transgene as much as their nuclear donor cells. In conclusions, (1) pig cloning can be carried out by TA or DA nuclear transfer methods, (2) expression of transgenes can be maintained to cloned pigs from the nuclear donor cells derived from transgenic animals.
authors.affiliation: Animal Engineering Research Institute, Midorigahara, Tsukuba, Ibaraki 300-2646, Japan. t.fujimura@nipponham.co.jp
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Fujimura
authors.forename: Mayuko
authors.initials: M
authors.lastname: Kurome
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Murakami
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Takahagi
authors.forename: Katsuyoshi
authors.initials: K
authors.lastname: Matsunami
authors.forename: Shinichi
authors.initials: S
authors.lastname: Shimanuki
authors.forename: Kohei
authors.initials: K
authors.lastname: Suzuki
authors.forename: Shuji
authors.initials: S
authors.lastname: Miyagawa
authors.forename: Ryota
authors.initials: R
authors.lastname: Shirakura
authors.forename: Tamotsu
authors.initials: T
authors.lastname: Shigehisa
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nagashima
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001755:Blastocyst / Q000166:cytology*; D018958:CD55 Antigens / Q000235:genetics*; D019976:Cloning, Organism / Q000379:methods*; D004624:Embryo Transfer; D005260:Female; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics*; D009865:Oocytes / Q000166:cytology*; D013552:Swine
chemical_list: D018958:CD55 Antigens; D017351:N-Acetylglucosaminyltransferases; C054599:beta-1,4-mannosyl-glycoprotein beta-1,4-N-acetylglucosaminyltransferase