公开(公告)号：KR1020050022416A

公开(公告)日：2005-03-08

申请号：KR1020030060612

申请日：2003-08-30

Applicant: 재단법인서울대학교산학협력재단

Inventor： 황우석 ,  이병천 ,  강성근 ,  현상환 ,  이갑상 ,  김혜수 ,  김수 ,  정연우 ,  남동현 ,  전현용 ,  염수청 ,  김순웅

IPC: C12N5/16

CPC classification number: C12N15/8778 ,  A01K67/0273 ,  A01K2227/108 ,  A01K2267/01 ,  A61D19/04 ,  C12N15/8509 ,  C12N2015/8518 ,  C12N2517/00

Abstract: PURPOSE: A transgenic cloned pig producing human erythropoietin and a method for producing the same transgenic cloned pig are provided, thereby producing a large quantity of human erythropoietin in the urinary bladder of the transgenic cloned pig with reduced costs. CONSTITUTION: The nuclear transfer embryo produced by a fusion of a pig-derived enucleated oocyte with a nucleus of a pig-derived somatic cell transfected and targeted with a gene encoding human erythropoietin is provided. The transgenic cloned pig expresses human erythropoietin in the urinary bladder of pig. The method for producing a transgenic cloned pig producing human erythropoietin comprises the steps of: (a) transfecting and targeting a gene encoding human erythropoietin into a pig-derived somatic cell line to prepare a nuclear donor cell; (b) removing cumulus cells surrounding a recipient oocyte and then the cytoplasm including the first polar body from the oocyte to prepare a pig-derived enucleated recipient oocyte; (c) transferring the transfected nuclear donor cell into the enucleated recipient oocyte and carrying out cell fusion to generate nuclear transfer embryos; and (d) transplanting the nuclear transfer embryo into a surrogate mother pig to produce live offspring.

Abstract translation: 目的：提供产生人促红细胞生成素的转基因克隆猪和产生相同转基因克隆猪的方法，从而以降低的成本在转基因克隆猪的膀胱中产生大量人促红细胞生成素。 构成：提供了通过将猪衍生的去核卵母细胞与转染并用编码人红细胞生成素的基因靶向的来源于体细胞的细胞核的融合产生的核转移胚。 转基因克隆猪在猪的膀胱中表达人促红细胞生成素。 生产人促红细胞生成素的转基因克隆猪的方法包括以下步骤：（a）将编码人促红细胞生成素的基因转染并靶向猪衍生的体细胞系以制备核供体细胞; （b）去除卵母细胞周围的卵丘细胞，然后从卵母细胞中除去包括第一极体的细胞质，以制备猪衍生的去核受体卵母细胞; （c）将转染的核供体细胞转移到去核的受体卵母细胞中并进行细胞融合以产生核转移胚胎; 和（d）将核转移胚胎移植到替代母猪中以产生活的后代。

公开(公告)号：KR1020040074108A

公开(公告)日：2004-08-21

申请号：KR1020047010333

申请日：2001-12-29

Applicant: 재단법인서울대학교산학협력재단

Inventor： 황우석 ,  이병천 ,  강성근 ,  한재용 ,  임정묵 ,  이창규 ,  이은송 ,  정의배 ,  조종기 ,  김대영 ,  현상환 ,  이갑상 ,  김혜수 ,  이소현 ,  이성철 ,  염수청

IPC: A01K67/02 ,  C12N5/07 ,  C12N15/63

Abstract: PURPOSE: A GFP(Green Fluorescent Protein)-transfected clone pig, a GT(alpha 1, 3 galactrosyltransferase) knock-out clone pig and a production method thereof are provided to produce a clone pig with a determined genetic character using transfection and targeting technologies. CONSTITUTION: The production method of the GFP-transfected clone pig comprises the steps of: preparing a cell line taken and cultured from a pig as a nucleus donating cell; introducing the GFP clone to the nucleus donating cell and expressing by forming a lipid(or a positive charge polymer)-DNA structure complex, adding the complex to the nucleus donating cell culture solution and culturing; producing a transfected embryo by transplanting the cell to an enucleated pig embryo and activating; and producing a piglet by transplanting the embryo to a surrogate mother.

Abstract translation: 目的：提供GFP（绿色荧光蛋白）转染的克隆猪，GT（α1,3半乳糖基转移酶）敲除克隆猪及其制备方法，以使用转染和靶向技术产生具有确定遗传特征的克隆猪 。 构成：GFP转染的克隆猪的制备方法包括以下步骤：制备从猪获得并培养为细胞核的细胞系; 将GFP克隆引入提供细胞核并通过形成脂质（或正电荷聚合物）-DNA结构复合物进行表达，将复合物加入给予细胞培养液的细胞核并培养; 通过将细胞移植到去核的猪胚胎并激活来产生转染的胚胎; 并通过将胚胎移植到替代母亲来生产仔猪。