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公开(公告)号：KR100500412B1
公开(公告)日：2005-07-12
申请号：KR1020030014132
申请日：2003-03-06
IPC: C12N5/16
Abstract: 본 발명은 동물의 복제란 제조방법에 관한 것으로서, 보다 상세하게는 난자의 핵을 염색하지 않고 제 1 극체와 10 내지 20%의 세포질을 흡입(suction)하여 난자의 핵을 제거하는 단계; 제 1 극체와 핵을 제거한 상기 난자의 위란강에 체세포를 주입하여 난자-체세포 복합체를 제조하는 단계; 및 ⅲ) 0.5 내지 2 ㎸/㎝, 6 내지 150 ㎲로 1 내지 3회 시행하는 전기적 자극으로 상기 난자-체세포 복합체를 세포융합 및 활성화시키는 단계를 포함하는 동물의 복제란 제조방법에 관한 것이다. 본 발명의 복제란 제조방법은 고효율의 제핵률을 나타낼 뿐만 아니라 복제과정을 단순화시킴으로써 다양한 생물의 체세포를 복제하는데 유용하게 사용될 수 있다.
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12.发明公开세포성 면역 억제를 위한 인간 ＦａｓＬ 단백질을 발현하는 형질전환 돼지 클론 체세포주 및 이를 이용한 형질전환 복제돼지 有权表达人体蛋白质以抑制细胞免疫的转基因克隆细胞系和使用其的转基因蛋白
公开(公告)号：KR1020110093132A
公开(公告)日：2011-08-18
申请号：KR1020100012983
申请日：2010-02-11
Applicant: (주)엠젠
IPC: C12N5/10 , C12N15/85 , C12N15/12 , A01K67/027
Abstract: PURPOSE: A transgenic porcine clonal somatic cell line which expresses human FasL gene in a tissue and cells is provided to reduce cytotoxicity and to minimize immune rejection. CONSTITUTION: A transgenic porcine clonal cell line contains human FasL gene expression vector and is deposited in the number KCLRF-BP-00194. A method for preparing the somatic cell line comprises: a step of isolating somatic cells from porcine fetus; a step of preparing hFasL gene expression vector and introducing to the somatic cells; and a step of selecting expression vector-introduced somatic cells and culturing.
Abstract translation: 目的：提供在组织和细胞中表达人FasL基因的转基因猪克隆体细胞系，以减少细胞毒性并使免疫排斥最小化。 构成：转基因猪克隆细胞系含有人FasL基因表达载体，并以KCLRF-BP-00194编号。 制备体细胞系的方法包括：从猪胎儿中分离体细胞的步骤; 制备hFasL基因表达载体并引入体细胞的步骤; 选择表达载体介导的体细胞和培养的步骤。
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公开(公告)号：KR1020090068646A
公开(公告)日：2009-06-29
申请号：KR1020070136343
申请日：2007-12-24
Applicant: (주)엠젠
Abstract: A method for producing transgenic pig which expresses HLA-G gene and DAF gene is provided to suppresses immune rejection of natural killer cell(NK cell) and use for cell theraphy and xeno-transplantation. A method for producing transgenic pig clone cell line which expresses human leukocyte antigen-G(HLA-G) gene and decay-accelerating factor(DAF) gene comprises: a step of isolating a somatic cell from HLA-G clonal pig; a step of inducing an expression vector containing DAF gene into the somatic cell; and a step of screening somatic cell in which the expression vector is induced. The expression vector is pCX-EGFP-BSD as a cloning vector. A method for producing the transgenic pig which expresses HLA-G gene and DAF gene comprises: a step of culturing the transgenic pig clone somatic cell line; a step of removing a nucleus of ovum and fusing clone somatic cell; and a step of transplanting the ovum to surrogate mother pig.
Abstract translation: 提供了表达HLA-G基因和DAF基因的转基因猪的制备方法，以抑制自然杀伤细胞（NK细胞）的免疫排斥，并用于细胞治疗和异种移植。 用于生产表达人白细胞抗原-G（HLA-G）基因和衰老加速因子（DAF））基因的转基因猪克隆细胞系的方法包括：从HLA-G克隆猪分离体细胞的步骤; 诱导含有DAF基因的表达载体进入体细胞的步骤; 以及筛选其中诱导表达载体的体细胞的步骤。 表达载体是作为克隆载体的pCX-EGFP-BSD。 一种生产表达HLA-G基因和DAF基因的转基因猪的方法包括：培养转基因猪克隆体细胞系的步骤; 去除卵的细胞核并融合克隆体细胞的步骤; 并将卵子移植到替代母猪的步骤。
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公开(公告)号：KR1020080112763A
公开(公告)日：2008-12-26
申请号：KR1020070061572
申请日：2007-06-22
Applicant: (주)엠젠 , 경남과학기술대학교 산학협력단
IPC: A01K67/027
CPC classification number: A01K67/0273 , A01K2217/052 , A01K2227/101 , A01K2267/0331 , C12N15/8509
Abstract: A manufacturing method of the transformed reproduction goat producing hgm-csf and transformation reproduction goat produced by the method are provided. A transformation reproduction goat manufacturing method for producing the CSF protein comprises: a step for separating the somatic cell from the fetus of goat or the female goat; a step for introducing into the somatic cell after preparing expression vector including the polynucleotide coding CSF; a step for cultivating after selecting the clone somatic cell in which the expression vector is introduced; a step for removing the nucleus of the ovum collected from the mother animal goat and fusing with the clone somatic cell; and a step for transplanting the fused clone egg into the goat.
Abstract translation: 提供了通过该方法生产的产生hgm-csf的转化繁殖山羊和转化繁殖山羊的制造方法。 用于产生CSF蛋白质的转化繁殖山羊制造方法包括：从山羊或雌性山羊胎儿分离体细胞的步骤; 在制备包括编码CSF的多核苷酸的表达载体之后引入体细胞的步骤; 选择其中导入了表达载体的克隆体细胞后进行培养的步骤; 去除从母体动物山羊收集的卵子的核并与克隆体细胞融合的步骤; 以及将融合的克隆蛋移植到山羊中的步骤。
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公开(公告)号：KR1020080034556A
公开(公告)日：2008-04-22
申请号：KR1020060100639
申请日：2006-10-17
Applicant: 경남과학기술대학교 산학협력단 , (주)엠젠
Abstract: A production method of a transgenic cloned embryo having a goat somatic cell nucleus transfected with hGM-CSF(human granulocyte macrophage-colony stimulating factor) gene is provided to produce a cloned goat expressing the hGM-CSF from the mammary gland, thereby improving the production yield of hGM-CSF. A production method of a transgenic cloned goat embryo comprises the steps of: (1) transforming a somatic cell of goat with a recombinant expression vector containing an hGM-CSF gene to prepare a nucleus donor cell; (2) collecting an ovum from the goat to prepare a nucleus recipient ovum; and (3) denucleating the recipient ovum, transplanting the nucleus donor cell into the recipient ovum and fusing them to prepare the transgenic cloned embryo.
Abstract translation: 提供了具有用hGM-CSF（人粒细胞巨噬细胞集落刺激因子）基因转染的山羊体细胞核的转基因克隆胚胎的制备方法，以从乳腺产生表达hGM-CSF的克隆山羊，从而提高产量 hGM-CSF产量。 转基因克隆山羊胚胎的制备方法包括以下步骤：（1）用含有hGM-CSF基因的重组表达载体转化山羊的体细胞以制备核供体细胞; （2）从山羊收集卵，制备受精卵; （3）使受体卵脱核，将核供体细胞移植到受体卵中并使其融合以制备转基因克隆的胚胎。
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Patent Agency Ranking
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公开(公告)号：KR1020070024069A
公开(公告)日：2007-03-02
申请号：KR1020050078585
申请日：2005-08-26
Abstract: A method for producing transgenic pig capable of expressing colony stimulating factor(CSF) gene in the mammary gland is provided to be able to easily obtain the transgenic pig, which is able to solve the problems of post-translational modification or mass-production and is used as a supplemental agent of treating leukemia and an immuno-potentiating agent, by expressing hGM-CSF in mammary gland. The method comprises the steps of: (a) isolating somatic cell from pig fetus on days 40-80 pregnancy; (b) after preparing an expression vector including a promoter specific to mammary gland such as alphas1-casein, beta-casein, beta-lactoglobulin and whey acidic protein; and a polynucleotide encoding the CSF such as hGM-CSF, introducing it into the somatic cell; (c) after selecting the expression vector-introduced clone somatic cell, culturing it; (d) removing nucleus of ovum collected from sow and fusing it with the clone somatic cell; and (e) transplanting the fused nuclear transfer embryos to surrogate mother pig and delivering piglets.
Abstract translation: 提供了能够在乳腺中表达集落刺激因子（CSF）基因的转基因猪的制备方法，以便能够容易地获得能够解决翻译后修饰或批量生产的问题的转基因猪，并且是 用作通过在乳腺中表达hGM-CSF来治疗白血病和免疫增强剂的补充剂。 该方法包括以下步骤：（a）在妊娠40-80天从胎儿分离体细胞; （b）在制备包括对乳腺特异性的启动子如α-α-酪蛋白，β-酪蛋白，β-乳球蛋白和乳清酸性蛋白质的表达载体后; 和编码CSF的多核苷酸，例如hGM-CSF，将其引入体细胞; （c）选择表达载体导入的克隆体细胞后进行培养; （d）去除从母猪收集的卵子的细胞核并与克隆体细胞融合; 和（e）将融合的核转移胚胎移植到替代母猪和递送仔猪。
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18.发明授权세포성 면역 억제를 위한 인간 ＦａｓＬ 단백질을 발현하는 형질전환 돼지 클론 체세포주 및 이를 이용한 형질전환 복제돼지 有权表达人FasL蛋白以抑制细胞免疫的转基因猪克隆体细胞系和使用其的转基因克隆猪
公开(公告)号：KR101147409B1
公开(公告)日：2012-05-22
申请号：KR1020100012983
申请日：2010-02-11
Applicant: (주)엠젠
IPC: C12N5/10 , C12N15/85 , C12N15/12 , A01K67/027
Abstract: PURPOSE: A transgenic porcine clonal somatic cell line which expresses human FasL gene in a tissue and cells is provided to reduce cytotoxicity and to minimize immune rejection. CONSTITUTION: A transgenic porcine clonal cell line contains human FasL gene expression vector and is deposited in the number KCLRF-BP-00194. A method for preparing the somatic cell line comprises: a step of isolating somatic cells from porcine fetus; a step of preparing hFasL gene expression vector and introducing to the somatic cells; and a step of selecting expression vector-introduced somatic cells and culturing.
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19.发明公开이종 이식시 일어나는 초급성 거부반응을 억제하기 위한ＣＤ５９ 유전자를 발현하는 형질전환 돼지 클론 체세포주및 그의 제조 방법 无效表达CD59基因的转基因小鼠皮肤细胞系抑制XENOTRANSPLANTATION所产生的高效抑制剂及其制备方法
公开(公告)号：KR1020080020251A
公开(公告)日：2008-03-05
申请号：KR1020060083489
申请日：2006-08-31
Applicant: (주)엠젠
Abstract: A transgenic pig clonal somatic cell line expressing protectin CD59 gene is provided to minimize complement activation by inhibiting cytotoxicity destruction in xenotransplantation, so that the cell line is useful for inhibition of hyperacute rejection occurred by xenotransplantation. A transgenic pig clonal somatic cell line expressing protectin CD59 gene(KCLRF-BP-00139) contains an expression cassette pcDNA-CD59(neo) encoding CD59 gene having the nucleotide sequence of SEQ ID NO:1. A method for manufacturing the transgenic pig clonal somatic cell line expressing protectin CD59 gene(KCLRF-BP-00139) comprises the steps of: (1) separating a somatic cell from the fetal pig by using primers of SEQ ID NOs:2 and 3; (2) preparing an expression vector containing a CD59 gene and transducing the expression vector into the somatic cell; and (3) selecting and culturing the transduced somatic cells. Further, the CD59 gene is SEQ ID NO:1.
Abstract translation: 提供表达保护素CD59基因的转基因猪克隆体细胞系以通过抑制异种移植中的细胞毒性破坏来最小化补体激活，使得细胞系可用于通过异种移植发生的超急性排斥的抑制。 表达保护素CD59基因（KCLRF-BP-00139）的转基因猪克隆体细胞系含有编码具有SEQ ID NO：1的核苷酸序列的CD59基因的表达盒pcDNA-CD59（neo）。 用于制备表达保护素CD59基因（KCLRF-BP-00139）的转基因猪克隆体细胞系的方法包括以下步骤：（1）使用SEQ ID NO：2和3引物从胎猪分离体细胞; （2）制备含有CD59基因并将表达载体转导入体细胞的表达载体; （3）选择和培养转导的体细胞。 此外，CD59基因是SEQ ID NO：1。
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公开(公告)号：KR100790514B1
公开(公告)日：2008-01-02
申请号：KR1020050078585
申请日：2005-08-26
Abstract: 본 발명은 인간 ＧＭ-ＣＳＦ를 유선에서 발현하는 형질전환 복제 돼지 및 그의 제조 방법에 관한 것이다. 보다 구체적으로는 1) 돼지 태아로부터 체세포를 분리하는 단계; 2) 유선(mammary gland) 특이적 프로모터 및 CSF(Colony stimulating factor, 세포집락 자극 인자)를 코딩하는 폴리뉴클레오티드를 포함하는 발현벡터를 제조한 후 상기 체세포에 도입시키는 단계; 3) 상기 발현 벡터가 도입된 클론 체세포를 선별한 후 배양하는 단계; 4) 모돈으로부터 채취한 난자의 핵을 제거하고 상기 체세포와 융합시키는 단계; 및 5) 상기 융합된 복제란을 대리모돈에 이식하고 자돈을 출산하는 단계를 포함하는 CSF 단백질을 발현하는 형질전환 돼지 및 그의 제조 방법에 관한 것이다. 본 발명의 제조방법에 의하여 제조된 형질전환 돼지는 기존 방법의 후-번역 수정(post-translational modification) 또는 대량 생산등의 문제점을 극복할 수 있으며, hGM-CSF를 유선에서 발현함으로써 쉽게 획득할 수 있는 잇점과 함께 이를 백혈병 치료 보조제 및 면역 강화제 등으로 사용될 수 있을 것으로 기대된다.
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