公开(公告)号：KR100984643B1

公开(公告)日：2010-10-01

申请号：KR1020070087391

申请日：2007-08-30

Applicant: 재단법인서울대학교산학협력재단

Inventor： 박재학 ,  김동재

IPC: C12N15/64 ,  C12N15/09 ,  C12N15/52

Abstract: 본 발명은 에스트로겐 반응성 리포터 유전자가 도입된 제브라피쉬 및 이를 이용한 에스트로겐 화합물의 스크리닝 방법에 관한 것이다. 보다 구체적으로, 본 발명은 제브라피쉬 뇌 아로마타제 유전자(zebrafish brain aromatase gene)의 인접 프로모터와 상기 프로모터에 의해 그 발현이 조절되는 형광단백질 리포터 유전자가 연결된 유전자 컨스트럭트가 도입된 제브라피쉬 및 여기에 시험물질을 처리한 후 리포터 유전자의 발현정도를 측정하는 것을 특징으로 하는 에스트로겐 화합물의 스크리닝 방법에 관한 것이다.
본 발명의 스크리닝 방법은 에스트로겐 활성을 가진 화합물을 결정하기 위한 새로운 생체 내(
in vivo ) 시스템으로서 상기 시스템은 생체 내 기능을 모사할 수 있으므로 생체 내에서 에스트로겐 활성을 가진 화합물을 신속하고 효과적으로 스크리닝할 수 있다.
에스트로겐 화합물, 제브라피쉬, 뇌 아로마타제 유전자, 스크리닝

公开(公告)号：KR1020090097503A

公开(公告)日：2009-09-16

申请号：KR1020080022666

申请日：2008-03-11

Applicant: 재단법인서울대학교산학협력재단

Inventor： 박재학 ,  석승혁

IPC: C12Q1/68

CPC classification number: C12Q1/6827 ,  C12Q2600/142

Abstract: A method for quantitatively analyzing the absolute value of fluorescence intensity and the amount of EGFP (Enhanced Green Fluorescent Protein) in a transgenic fish is provided to quantitatively measure the harmfulness of toxic material. A method for qualitatively analyzing the amount of GFP or EGFP of transgenic fish comprises: a first step of measuring the fluorescence intensity based on the amount of the GFP and purified GFP and forming standard linear equation; a second step of inserting a toxic material reactive gene promoter in a vector which expresses GFP or EGFP to produce the transgenic fish; a third step of measuring the fluorescence intensity which the transgenic fish expresses; a fourth step of calculating the amount of GFP or EGFP corresponding to the fluorescence intensity; and a step of measuring the amount of purified GFP or EGFP and forming standard linear equation.

Abstract translation: 提供了定量分析转基因鱼的荧光强度绝对值和EGFP（增强型绿色荧光蛋白）量的方法，以定量测定有毒物质的有害性。 用于定性分析转基因鱼的GFP或EGFP的量的方法包括：基于GFP和纯化的GFP的量测量荧光强度并形成标准线性方程的第一步骤; 将毒性物质反应性基因启动子插入到表达GFP或EGFP以产生转基因鱼的载体中的第二步骤; 测量转基因鱼表达的荧光强度的第三步; 计算对应于荧光强度的GFP或EGFP的量的第四步骤; 以及测量纯化的GFP或EGFP的量并形成标准线性方程的步骤。

公开(公告)号：KR1020080012449A

公开(公告)日：2008-02-12

申请号：KR1020060073275

申请日：2006-08-03

Applicant: 재단법인서울대학교산학협력재단

Inventor： 박재학

IPC: G01N33/53 ,  G01N33/50 ,  G01N33/68 ,  A61K39/00

Abstract: Diagnostic methods for SARS(severe acute respiratory syndrome) by using nucleocapside or spike protein are provided to improve the safety and accuracy of diagnosis and allow early diagnosis, so that they are applicable to general laboratories and hospitals without a risk of infection. A method for diagnosis of SARS comprises the steps of: amplifying a gene encoding SARS-CoV-N(SARS coronavirus-nucleocapsid protein) and amplifying the amplified gene to Escherichia coli to produce an SARS-CoV-N antigen; producing an antibody against the SARS-CoV-N antigen; and reacting a specimen containing the SARS-CoV-N antibody coupled with a coloring material such as enzyme or gold and the SARS-CoV antigen with a membrane adsorbing the SARS-CoV-N antibody. A diagnostic kit contains the membrane adsorbing the SARS-CoV-N antibody and solution containing the SARS-CoV-N antibody coupled with a coloring material. A method for preparing SARS-CoV-S(SARS coronavirus-spike protein) antigen comprises the steps of: amplifying a gene encoding SARS-CoV-S antigen and inserting the amplified gene into a vector; transfecting 293 T cell with an expression vector containing SARS-CoV-S antigen; and infecting the transformed 293 T cell with VSV(vesicular stomatitis vesicle) to produce SARS-CoV-S antigen. Further, the antibody is a single clone antibody.

Abstract translation: 提供了通过使用核壳蛋白或尖峰蛋白的SARS（严重急性呼吸综合征）的诊断方法，以提高诊断的安全性和准确性，并允许早期诊断，从而适用于没有感染风险的一般实验室和医院。 SARS诊断方法包括以下步骤：将编码SARS-CoV-N（SARS冠状病毒 - 核壳蛋白）的基因扩增，并将扩增的基因扩增至大肠杆菌以产生SARS-CoV-N抗原; 产生针对SARS-CoV-N抗原的抗体; 并将含有SARS-CoV-N抗体与着色材料如酶或金和SARS-CoV抗原偶联的样品与吸附SARS-CoV-N抗体的膜反应。 诊断试剂盒含有吸附SARS-CoV-N抗体的膜和含有与着色材料偶联的SARS-CoV-N抗体的溶液。 制备SARS-CoV-S（SARS冠状病毒 - 刺突蛋白）抗原的方法包括以下步骤：扩增编码SARS-CoV-S抗原的基因并将扩增的基因插入载体; 用含有SARS-CoV-S抗原的表达载体转染293T细胞; 并用VSV（水泡性小泡）感染转化的293T细胞以产生SARS-CoV-S抗原。 此外，抗体是单克隆抗体。

公开(公告)号：KR1020090097504A

公开(公告)日：2009-09-16

申请号：KR1020080022667

申请日：2008-03-11

Applicant: 재단법인서울대학교산학협력재단

Inventor： 박재학 ,  석승혁

IPC: C12Q1/68 ,  G06F19/28

CPC classification number: G01N33/5014 ,  C12Q1/6827 ,  C12Q2600/142

Abstract: A method for evaluating toxicity through the expression of fluorescence protein in a transgenic embryo of animal is provided to compare toxic degree between different species and analyze at the initial developing stage. A method for evaluating the effect on a toxic material in developing embryo comprises: a first step of injecting a fluorescence protein expression vector in which toxic material-reactive gene promoter is inserted in an embryo to produce a transgenic embryo of animal; and a second step of evaluating the expression of fluorescence protein by exposing the embryo with the toxic material. The animal is zebrafish. The used toxic material is arsenic.

Abstract translation: 提供通过动物转基因胚胎中荧光蛋白表达来评估毒性的方法，以比较不同物种之间的毒性程度，并在初始发育阶段进行分析。 用于评价对发育中的胚胎中的有毒物质的影响的方法包括：将荧光蛋白质表达载体注入第一步骤，其中将有毒物质反应性基因启动子插入胚胎中以产生动物的转基因胚胎; 以及通过用有毒物质暴露胚胎来评估荧光蛋白的表达的第二步骤。 动物是斑马鱼。 使用过的有毒物质是砷。

公开(公告)号：KR1020090022231A

公开(公告)日：2009-03-04

申请号：KR1020070087391

申请日：2007-08-30

Applicant: 재단법인서울대학교산학협력재단

Inventor： 박재학 ,  김동재

IPC: C12N15/64 ,  C12N15/09 ,  C12N15/52

Abstract: A zebrafish having an estrogen reactivity reporter gene and a screening method thereby are provided to screen compounds with estrogen activity in a living body. A recombinant plasmid introduced in a zebrafish fertilized egg comprises a gene construct for connecting an adjacent promoter of a zebrafish brain aromatase gene to a reporter gene whose expression is controlled by the promoter. The zebrafish brain aromatase gene is indicated as a sequence number 1. The report gene is selected from the group consisting of EGFP(enhanced green fluorescent protein), GFP(green fluorescence protein), RFP(red fluorescence protein), luciferase gene, and a beta- galactosidase gene.

Abstract translation: 提供具有雌激素反应性报告基因的斑马鱼及其筛选方法，以筛选活体内具有雌激素活性的化合物。 在斑马鱼受精卵中引入的重组质粒包括用于将斑马鱼脑芳香酶基因的相邻启动子连接到表达受启动子控制的报道基因的基因构建体。 报告基因选自EGFP（增强型绿色荧光蛋白），GFP（绿色荧光蛋白），RFP（红色荧光蛋白），萤光素酶基因，以及荧光素酶基因 β-半乳糖苷酶基因。

公开(公告)号：KR1020080009785A

公开(公告)日：2008-01-30

申请号：KR1020060069426

申请日：2006-07-25

Applicant: 재단법인서울대학교산학협력재단

Inventor： 박재학 ,  석승혁

IPC: C12N15/06 ,  C12N15/00

Abstract: A transgenic zebrafish for searching environmental contaminants and a method for preparing the same are provided to evaluate the preclinical effect and sensitivity by providing a biomarker for risk of the environmental contaminants. A method for preparing a transgenic zebrafish for searching environmental contaminants comprises the steps of: (a) cloning a pollutant response gene promoter of human such as HSRE(heat shock responsive element) and AhRE(aryl-hydrocarbon responsive element); (b) inserting the promoter into a vector expressing a fluorescent protein; and (c) introducing the vector into zebrafish. A transgenic zebrafish for searching environmental contaminants prepared by the method is characterized in that it radiates when it is exposed to environmental stress such as heat, cold, oxygen deficiency, Cd, L-azetidine-2-carboxylic acid, 2,3,7,8-tetrachlorodibenzo-p-dioxin, 3-methylcholanthrene, 3,4,5,3',4',5'-hexabromobiphenyl, Aroclor 1254 and benzo[a]pyrene.

Abstract translation: 提供了一种用于检索环境污染物的转基因斑马鱼及其制备方法，通过为环境污染物的风险提供生物标志物来评估临床前效应和敏感性。 制备用于检索环境污染物的转基因斑马鱼的方法包括以下步骤：（a）克隆人的污染物应答基因启动子如HSRE（热休克反应元件）和AhRE（芳烃响应元件）; （b）将启动子插入表达荧光蛋白的载体中; 和（c）将载体引入斑马鱼。 用于搜索通过该方法制备的环境污染物的转基因斑马鱼的特征在于当其暴露于环境胁迫如热，冷，缺氧，Cd，L-氮杂环丁烷-2-羧酸，2,3,7， 8-四氯二苯并对二恶英，3-甲基胆蒽，3,4,5,3'，4'，5'-六溴代二苯，Aroclor 1254和苯并[a]芘。