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    • 9. 发明公开
    • 곤충세포 발현 재조합 3ABC 비구조단백질 항원 및 단클론항체를 이용한 구제역 진단방법
    • 使用在INSEC细胞和单克隆抗体中表达的重组蛋白3ABC非结构蛋白的足球疾病的诊断方法
    • KR1020030052858A
    • 2003-06-27
    • KR1020010082976
    • 2001-12-21
    • 대한민국(농림축산식품부 농림축산검역본부장)
    • 권창희현방훈고영준이광녕구복경엄재구나진주남향미박지용계수정김인중이세영최광석손현주주이석
    • C12N15/33
    • C12N15/866C07K14/005C07K16/1009C07K2317/14C12N5/163C12N2770/32131G01N33/56983G01N33/577G01N2333/09
    • PURPOSE: Diagnostic methods of foot-and-mouth disease using recombinant 3ABC non-structural protein expressed in insect cells and a monoclonal antibody is provided, thereby more rapidly and accurately diagnosing the foot-and-mouth disease than the prior methods. CONSTITUTION: A gene encoding a recombinant 3ABC non-structural protein derived from Korean foot-and-mouth disease virus O/SKR/2000 has the nucleotide sequence of SEQ ID NO: 1. A recombinant baculovirus to be expressed in insect cells is prepared by co-transfection with a recombinant vector containing the recombinant 3ABC non-structural protein gene of SEQ ID NO: 1. The recombinant 3ABC non-structural protein is expressed by infection of the insect cells with the recombinant baculovirus. A hybridoma cell line 3F-11(KCTC 10138BP) is prepared by introducing the recombinant 3ABC non-structural protein expressed in E. coli into an animal, collecting an immunized cell from the animal and fusing the immunized cell with a cancer cell. A monoclonal antibody is produced from the hybridoma cell line 3F-11(KCTC 10138BP). A diagnostic method of foot-and-mouth disease comprises the steps of: (1) diluting the recombinant 3ABC non-structural protein in a coating buffer solution and pouring the diluate on a plate; (2) reacting the testing serum with the plate; (3) reacting a conjugate, which binds with an antibody for foot-and-mouth disease virus in the testing serum and has an enzyme, a radioactive material or a fluorescent material, with the testing serum; and (4) measuring intensity of the enzyme reaction, fluorescence reaction or radiation reaction with the conjugate.
    • 目的:提供使用在昆虫细胞中表达的重组3ABC非结构蛋白和单克隆抗体的口蹄疫诊断方法,从而比现有方法更快速,准确地诊断口蹄疫。 构成:编码来自韩国口蹄疫病毒O / SKR / 2000的重组体3ABC非结构蛋白的基​​因具有SEQ ID NO:1的核苷酸序列。在昆虫细胞中表达的重组杆状病毒通过 用含有SEQ ID NO:1的重组3ABC非结构蛋白基因的重组载体共转染。重组体3ABC非结构蛋白用重组杆状病毒感染昆虫细胞表达。 通过将在大肠杆菌中表达的重组3ABC非结构蛋白质导入动物中,从动物中收集免疫的细胞并将免疫的细胞与癌细胞融合来制备杂交瘤细胞系3F-11(KCTC 10138BP)。 从杂交瘤细胞系3F-11(KCTC 10138BP)产生单克隆抗体。 口蹄疫的诊断方法包括以下步骤:(1)在包被缓冲溶液中稀释重组体3ABC非结构蛋白,将稀释液倒入平板上; (2)使测试血清与板反应; (3)使与测试血清中的口蹄疫病毒抗体结合并具有酶,放射性物质或荧光材料的缀合物与测试血清反应; 和(4)测量酶反应的强度,荧光反应或与缀合物的辐射反应。