KR960031610A - Tobacco Plants Producing Assembled Antibodies Against Hepatitis B Virus - Google Patents

Tobacco Plants Producing Assembled Antibodies Against Hepatitis B Virus Download PDF

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KR960031610A
KR960031610A KR1019950002371A KR19950002371A KR960031610A KR 960031610 A KR960031610 A KR 960031610A KR 1019950002371 A KR1019950002371 A KR 1019950002371A KR 19950002371 A KR19950002371 A KR 19950002371A KR 960031610 A KR960031610 A KR 960031610A
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hepatitis
tobacco
tobacco plant
recombinant vector
virus antibody
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KR0155453B1 (en
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이호설
정창호
김신제
백경희
권석윤
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김홍기
금호석유화학 주식회사
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
    • C07K16/081—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from DNA viruses
    • C07K16/082—Hepadnaviridae, e.g. hepatitis B virus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
    • C12N15/8257—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits for the production of primary gene products, e.g. pharmaceutical products, interferon

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Abstract

본 발명은 B형 간염바이러스의 항체를 담배식물(Nicotiana tabacum)로부터 생산하는 과정에서 사용되며 B형 간염바이러스 항체의 헤비체인 또는 카파체인 유전자를 각각 함유하는 재조합 벡터, 재조합 벡터로 미생물을 형질전환시켜 제조된 균주, 상기 균주를 매개로 하여 항체의 헤비체인 또는 카파체인 유전자로 각각 형질전환된 담배식물, 상기 담배식물로부터 제조된 헤비체인 및 카파체인 단백질, 상기 담배식물을 상호 교배(Cross Pollenation)하여 헤비체인 유전자 및 카파체인 유전자가 한 개체내에 도입된 담배식물, 각각 상이한 개체의 담배식물로부터 얻어진 헤비체인 및 카파체인 단백질을 결합시키거나 상호 교배에 의해 동물의 면역체계에서 존재하는 조립된 항체를 생산할 수 있도록 형질전환된 담배식물로부터 제조되는 B형 간염바이러스의 조립된 항체, 및 이러한 B형 간염바이러스의 조립된 항체를 유효 성분으로 함유하는 B형 간염 및 간암 치료체에 관한 것이다.The present invention is used in the process of producing the antibody of hepatitis B virus from tobacco plants (Nicotiana tabacum) and transforming microorganisms with recombinant vector, recombinant vector containing heavy chain or kappa chain gene of hepatitis B virus antibody, respectively. To prepare a strain, a tobacco plant transformed with the heavy chain or kappa chain gene of the antibody through the strain, heavy chain and kappa chain protein prepared from the tobacco plant, cross pollen cross-linking (tobacco plant) Heavy chain genes and kappachain genes can be used to produce assembled antibodies present in the animal's immune system by combining or crossbreeding tobacco plants introduced into one individual, heavy chain and kappachain proteins obtained from tobacco plants of different individuals, respectively. Assembled Hepatitis B Virus Prepared from Tobacco Plants An antibody and a hepatitis B and liver cancer therapeutic agent containing an assembled antibody of such hepatitis B virus as an active ingredient.

Description

B형 간염바이러스에 대한 조립된 항체를 생산하는 담배식물Tobacco Plants Producing Assembled Antibodies Against Hepatitis B Virus

본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음Since this is an open matter, no full text was included.

제1도는 B형 간염바이러스의 Pre-S2 항원에 대한 항체를 구성하는 헤비체인 및 카파체인 유전자를 각각 플라스미드 pBKS-1에 삽입함으로써 재조합 플라스미드 pBHC-1 및 pBKC-1를 제조하는 과정을 도시한 것이다,FIG. 1 shows the preparation of recombinant plasmids pBHC-1 and pBKC-1 by inserting the heavy chain and kappa chain genes constituting the antibody against the Pre-S2 antigen of hepatitis B virus into plasmid pBKS-1, respectively. ,

제2도는 B형 간염바이러스의 Pre-S2 항원에 대한 항체를 구성하는 헤비체인 및 카파체인 유전자가 플라스미드 pBKS-1에 삽입되었음을 확인하기 위한 자외선 촬영도이다,Figure 2 is an ultraviolet photograph to confirm that the heavy chain and kappa chain genes constituting the antibody to the Pre-S2 antigen of hepatitis B virus inserted into the plasmid pBKS-1,

제3도는 형질전환된 담배의 노던(Northern)분석 결과를 나타낸 것이다.Figure 3 shows the results of the Northern (Northern) analysis of the transformed tobacco.

Claims (23)

B형 간염바이러스 항체의 헤비체인 유전자를 고등식물 형질전환용 벡터에 삽입시켜 제조된 재조합 벡터.A recombinant vector prepared by inserting a heavy chain gene of hepatitis B virus antibody into a vector for transformation of higher plants. 제1항에 있어서, 형질전환용 벡터가 pBKS-1인 재조합 벡터.The recombinant vector of claim 1, wherein the transformation vector is pBKS-1. 제1항에 있어서, pBHC-1인 재조합 벡터.The recombinant vector of claim 1, wherein the recombinant vector is pBHC-1. B형 간염바이러스 항체의 헤비체인 유전자를 고등식물 형질전화용 벡터에 삽입시켜 제조된 재조합 벡터로 미생물을 형질전환시켜 제조된 균주.A strain prepared by transforming a microorganism with a recombinant vector prepared by inserting a heavy chain gene of a hepatitis B virus antibody into a vector for transforming a higher plant. 제4항에 있어서, 미생물로서 아그로박테리움 튜머페시언스(Agrobacterium tumefaciens)를 사용함을 특징으로 하는 균주.The strain according to claim 4, wherein Agrobacterium tumefaciens is used as the microorganism. 제5항에 있어서, 미생물이 아그로박테리움 튜머페시언스 LBA4404임을 특징으로 하는 균주.6. The strain of claim 5, wherein the microorganism is Agrobacterium tumer Pessence LBA4404. B형 간염바이러스 항체의 헤비체인 유전자를 고등식물 형질전환용 벡터에 삽입시켜 제조된 재조합 벡터로 형질전환시킨 균주에 의해 형질전환된 담배식물.A tobacco plant transformed by a strain transformed with a recombinant vector prepared by inserting a heavy chain gene of hepatitis B virus antibody into a higher plant transformation vector. 제7항에 있어서, 담배종자, 담배조직, 전체 담배식물의 각 형태를 포함하는 담배식물.8. A tobacco plant according to claim 7, comprising each type of tobacco seed, tobacco tissue, whole tobacco plant. 제7항의 담배식물로부터 분리 정제한 B형 간염바이러스 항체의 헤비체인 단백질.A heavy chain protein of hepatitis B virus antibody isolated and purified from the tobacco plant of claim 7. B형 간염바이러스 항체의 카파체인 유전자를 고등식물 형질전환용 벡터에 삽입시켜 제조된 재조합 벡터.A recombinant vector prepared by inserting a kappa chain gene of hepatitis B virus antibody into a vector for transformation of higher plants. 제10항에 있어서, 형질전환용 벡터가 pBKS-1인 재조합 벡터.The recombinant vector of claim 10, wherein the transformation vector is pBKS-1. 제10항에 있어서, pBKC-1인 재조합 벡터.The recombinant vector of claim 10, wherein said recombinant vector is pBKC-1. B형 간염바이러스 항체의 카파체인 유전자를 고등식물 형질전환용 벡터에 삽입시켜 제조된 재조합 벡터로 미생물을 형질전환시켜 제조된 균주.A strain prepared by transforming a microorganism with a recombinant vector prepared by inserting a kappa chain gene of hepatitis B virus antibody into a higher plant transformation vector. 제13항에 있어서, 미생물로서 아그로박테리움 튜머페시언스(Agrobacterium tumefaciens)를 사용함을 특징으로 하는 균주.(시켜 카파체인 유전자와 헤비체인 유전자가 도입된 담배식물)A strain according to claim 13, characterized in that it uses Agrobacterium tumefaciens as a microorganism (tobacco plants into which the kappachain gene and the heavychain gene are introduced). 제14항에 있어서, 미생물이 아그로박테리움 튜머페시언스 LBA4404임을 특징으로 하는 균주.15. The strain according to claim 14, wherein the microorganism is Agrobacterium tumerpesis LBA4404. B형 간염바이러스 항체의 카파체인 유전자를 고등식물 형질전환용 벡터에 삽입시켜 제조된 재조합 벡터로 형질전환시킨 균주에 의해 형질전환된 담배식물.A tobacco plant transformed by a strain transformed with a recombinant vector prepared by inserting a kappa chain gene of a hepatitis B virus antibody into a vector for transforming a higher plant. 제16항에 있어서, 담배종자, 담배조직, 전체 담배식물의 각 형태를 포함하는 담배식물.17. The tobacco plant of claim 16, comprising each form of tobacco seed, tobacco tissue, whole tobacco plant. 제16항의 담배식물로부터 분리 정제한 B형 간염바이러스 항체의 카파체인 단백질.A kappache protein of hepatitis B virus antibody purified from the tobacco plant of claim 16. 제7항의 담배식물과 제16항의 담배식물을 상호 교배(Cross Pollenation)시켜 카파체인 유전자와 헤비체인 유전자가 도입된 담배식물.A tobacco plant in which a kappa chain gene and a heavy chain gene are introduced by cross-crossing the tobacco plant of claim 7 and the tobacco plant of claim 16. 제19항에 있어서, 담배종자, 담배조직, 전체 담배식물의 각 형태를 포함하는 담배식물.20. Tobacco plant according to claim 19, comprising each form of tobacco seed, tobacco tissue, whole tobacco plant. 제7항의 담배식물을 재배하여 이루부터 B형 간염바이러스 항체의 헤비체인 단백질을 분리 및 정제하고, 제16항의 담배식물을 재배하여 이로부터 B형 간염바이러스 항체의 카파체인 단백질을 분리 및 정제한 다음 두가지 단백질을 결합시킴으로써 제조됨을 특징으로 하는 조립된 B형 간염바이러스 항체.After cultivating the tobacco plant of claim 7 to isolate and purify the heavy chain protein of hepatitis B virus antibody, and to grow the tobacco plant of claim 16 to isolate and purify the kappa chain protein of hepatitis B virus antibody from An assembled hepatitis B virus antibody, characterized in that it is prepared by combining two proteins. 제19항의 담배식물을 재배하고 이로부터 항체 단백질을 분리 및 정제하여 제조되는 조립된 B형 간염바이러스 항체.An assembled hepatitis B virus antibody prepared by cultivating the tobacco plant of claim 19 and isolating and purifying the antibody protein therefrom. 제21항 또는 22항의 조립된 B형 간염바이러스 항체를 유효 성분으로 함유하는 B형 간염 또는 간암 치료제.A therapeutic agent for hepatitis B or liver cancer containing the assembled hepatitis B virus antibody of claim 21 or 22 as an active ingredient. ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.※ Note: The disclosure is based on the initial application.
KR1019950002371A 1995-02-09 1995-02-09 Method of combined antibody against hepatitis-b virus by nicotiana tabacum KR0155453B1 (en)

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