Abstract:
본 발명은 치료용 단백질의 대량 생산을 위하여, 5' 아암(arm)으로서, 서열번호 1로 기재되는 핵산서열을 갖는 폴리뉴클레오티드로 구성된 소 베타-카제인 게놈 DNA의 5176 bp 프로모터, 엑손 1 및 2를 포함하는 5' 말단 단편; 치료용 단백질(therapeutic protein)을 암호화하는 유전자; 및 3' 아암으로서, 서열번호 2로 기재되는 핵산서열을 갖는 폴리뉴클레오티드로 구성된 소 베타-카제인 게놈 DNA의 엑손 3 및 엑손 4를 포함하는, 치료용 단백질 생산용 넉-인(knock in) 벡터를 제공한다.
Abstract:
PURPOSE: A method for producing porcine somatic cells for producing a human basic FGF(hFGF) with a knock-in vector is provided to express the hFGF with high concentration and to ensure stability and excellent physical activity. CONSTITUTION: A method for manufacturing porcine somatic cells in which porcine beta-casein is targeted comprises: a step of transforming porcine somatic cells with porcine beta-casein gene targeting knock-in vector ; a step of culturing the porcine somatic cells to induce homology recombination; and a step of selecting porcine somatic cells in which porcine beta-casein gene is targeted by the homology recombination. A method for transforming porcine comprises: a step of transforming porcine somatic cells with porcine beta-casein gene targeting knock-in vector; a step of culturing porcine somatic cells to induce homology recombination; a step of selecting the porcine somatic cells; a step of removing nucleus of the porcine ovary and introducing the targeted cells to obtain a fertilized egg in which somatic cells are nuclear transplanted; and a step of implanting the fertilized egg.
Abstract:
A knock-in vector for producing a bioactive substance is provided to efficiently and stably express bFGF (basic Fibroblast Growth Factor) and GFP (Green Fluorescent Protein) from porcine milk. A knock-in vector for producing a bioactive substance comprises 5' end fragment, gene coding the bioactive substance, and 3' end fragment. The 5' end fragment is 5' arm and comprises 2.65kb of promoter, one axon and one intron. The 3' end fragment is 3' arm, and comprises an axon 5, 6, and 7 of porcine beta-casein genome DNA. A nuclear localization signal (NLS) is additionally connected to the 5' end. The knock-in vector comprises a positive selectable marker and negative selectable marker. The bioactive substance is basic fibroblast growth factor (bFGF) or green fluorescent protein (GFP).
Abstract:
The present invention provides a transgenic animal produced by using a recombinant expression vector which includes: a promoter for inducing a fat cell specific expression for researching a function of PEN2 genes in obesity and metabolic syndrome; and presenilin enhancer 2 genes connected to the promotor to be operated.