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:
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:
본 발명은 CSF(Colony stimulating factor, 세포집락 자극 인자), 바람직하게는 사람의 과립구 대식세포-콜로니 자극 인자(hGM-CSF)를 유선에서 생산하는 형질전환 복제 산양 및 그의 제조 방법에 관한 것으로서, 보다 구체적으로는 1) 산양 의 태아 또는 암컷 산양으로부터 체세포를 분리하는 단계; 2) 유선(mammary gland) 특이적 프로모터 및 CSF(Colony stimulating factor, 세포집락 자극 인자)유전자를 코딩하는 폴리뉴클레오티드를 포함하는 발현벡터를 제조한 후 상기 체세포에 도입시키는 단계; 3) 상기 발현 벡터가 도입된 클론 체세포를 선별한 후 배양하는 단계; 4) 어미 산양으로부터 채취한 난자의 핵을 제거하고 상기 클론 체세포와 융합시키는 단계; 및 5) 상기 융합된 복제란을 수란 산양(대리모)에 이식하여 산자를 출산하는 단계를 포함하는 CSF 단백질을 생산하는 형질전환 복제 산양 및 그의 제조 방법에 관한 것이다. 본 발명의 제조방법에 의하여 제조된 형질전환 복제 산양은 기존 방법의 후-번역수정(post-translational modification) 또는 대량 생산 등의 문제점을 극복할 수 있으며, CSF 단백질을 유선에서 생산함으로써 쉽게 획득할 수 있는 이점과 함께 이를 백혈병 치료 보조제 및 면역 강화제 등으로 사용될 수 있을 것으로 기대된다. hGM-CSF, 형질전환된 복제 산양
Abstract:
A transformed porcine clonal somatic cell line which expresses an US6 protein is provided to suppress the MHC class I expression, minimize the immunerejection and use for the xeno-transplantation. A transformed porcine clonal somatic cell line(deposit number: KCLRF-BP-00158) comprises an US6(unique short 6) gene expression vector of cytomegalovirus(CMV), pcDNA3.1/myc-His(-B). A method for manufacturing the transformed porcine clonal somatic cell line comprises: a step of isolating the somatic cell from a pig fetus; a step of introducing the pcDNA3.1/myc-His(-B) of cytomegalovirus into the somatic cell; and a step of screening and culturing the somatic cell having the expression vector. The pig fetus is 20-50 days old.
Abstract:
본 발명은 US2 단백질을 발현하는 형질전환 돼지 클론 체세포주 및 상기 세포주의 제조 방법에 관한 것으로서, 보다 상세하게는 1) 돼지 태아로부터 체세포를 분리하는 단계; 2) US2 유전자를 포함하는 발현벡터를 제조한 후 단계 1의 체세포에 도입시키는 단계; 및 3) 상기 발현벡터가 도입된 체세포를 선별한 후 배양하는 단계를 포함하는 US2 단백질을 발현하는 형질전환 돼지 클론 체세포주의 제조 방법에 관한 것이다. 본 발명의 제조 방법에 의한 US2 단백질을 발현하는 돼지 클론 체세포주는 주요 조직적합성 복합체 제 1형 항원(major histocompatibility complex: MHC Class I)의 발현이 저해되어 세포성 면역거부반응이 최소화됨으로써, 핵이식을 통한 US2 단백질 발현 돼지의 생산에 이용하여 이종장기이식 연구 및 이의 산업적 이용성에 유용하게 사용할 수 있다.
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:
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:
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:
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.