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:
Provided is a method of producing a transgenic pig expressing a green-fluorescent protein to be effectively used in a field of producing a transgenic protein for curing. The method of producing a transgenic pig expressing a green-fluorescent protein comprises the steps of (1) separating somatic cells from a pig fetus, (2) producing a expression vector pCX-EGFP/neo with a fluorescent protein gene and then introducing the somatic cells of the (1) step thereinto, (3) selecting clone somatic cells introduced with the expression vector pCX-EGFP/neo and then cultivating the clone somatic cells, (4) removing a nucleus of an ovum taken from a mother pig and fusing it with the somatic cells, (5) transplanting the fused cloning egg in a surrogate mother pig and giving birth to a baby pig.
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 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:
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.