公开(公告)号：KR1020020040494A

公开(公告)日：2002-05-30

申请号：KR1020000070557

申请日：2000-11-24

Applicant: 재단법인서울대학교산학협력재단

Inventor： 김영식 ,  이진선 ,  문기영 ,  한범수 ,  안광석

IPC: C12N15/65

CPC classification number: C12N15/65 ,  C12Q2600/158

Abstract: PURPOSE: Provided are stable system for quantitative determination of NF-kappa B expression in human skin cells and its use in screening skin whitening agents, anti-inflammatory agents or anti-oxidants. The analytical system uses fluorescent substrate to make it very sensitive and simple and is used effectively in screening skin whitening agents, anti-inflammatory agents or anti-oxidants in natural products and organic compounds. CONSTITUTION: The determination system comprises: a fused expression vector, pNF-k B-SEAP-NPT from fusing a gene expressing NF-kappa B transcription reporter, selected from SEAP(secretory alkaline phosphatase), luciferase, beta-galactosidase and GFP(green fluorescent protein), with a selectable marker expression gene chosen from NPT(neomycin phosphotransferase), CAT(chloramphenicol acetyltransferase) and beta-galactosidase, lacZ; transfecting it to skin cell line; and quantitatively determining the expression of NF-kappa B from the above transfected skin cell line(KCLRF-BP-00035) by fluorometry.

Abstract translation: 目的：提供人体皮肤细胞中NF-κB表达定量测定的稳定系统及其在筛选皮肤美白剂，抗炎剂或抗氧化剂中的应用。 分析系统使用荧光底物使其非常灵敏和简单，并且有效地用于筛选天然产物和有机化合物中的皮肤增白剂，抗炎剂或抗氧化剂。 构成：测定系统包括：融合表达载体，pNF-k B-SEAP-NPT，其融合表达NF-κB转录报告基因的基因，其选自SEAP（分泌性碱性磷酸酶），荧光素酶，β-半乳糖苷酶和GFP（绿色 荧光蛋白），具有选自NPT（新霉素磷酸转移酶），CAT（氯霉素乙酰转移酶）和β-半乳糖苷酶，lacZ的选择性标记表达基因; 将其转染至皮肤细胞系; 并通过荧光测定定量测定来自上述转染的皮肤细胞系（KCLRF-BP-00035）的NF-κB的表达。

公开(公告)号：KR100414781B1

公开(公告)日：2004-01-13

申请号：KR1020000070557

申请日：2000-11-24

Applicant: 재단법인서울대학교산학협력재단

Inventor： 김영식 ,  이진선 ,  문기영 ,  한범수 ,  안광석

IPC: C12N15/65

Abstract: PURPOSE: Provided are stable system for quantitative determination of NF-kappa B expression in human skin cells and its use in screening skin whitening agents, anti-inflammatory agents or anti-oxidants. The analytical system uses fluorescent substrate to make it very sensitive and simple and is used effectively in screening skin whitening agents, anti-inflammatory agents or anti-oxidants in natural products and organic compounds. CONSTITUTION: The determination system comprises: a fused expression vector, pNF-k B-SEAP-NPT from fusing a gene expressing NF-kappa B transcription reporter, selected from SEAP(secretory alkaline phosphatase), luciferase, beta-galactosidase and GFP(green fluorescent protein), with a selectable marker expression gene chosen from NPT(neomycin phosphotransferase), CAT(chloramphenicol acetyltransferase) and beta-galactosidase, lacZ; transfecting it to skin cell line; and quantitatively determining the expression of NF-kappa B from the above transfected skin cell line(KCLRF-BP-00035) by fluorometry.

Abstract translation: 目的：提供用于定量测定人皮肤细胞中NF-κB表达的稳定系统及其用于筛选皮肤增白剂，抗炎剂或抗氧化剂的用途。 该分析系统使用荧光底物使其非常敏感和简单，并有效地用于筛选天然产品和有机化合物中的皮肤增白剂，抗炎剂或抗氧化剂。 组成：测定系统包括：融合表达NF-κB转录报道基因的融合表达载体pNF-k B-SEAP-NPT，选自SEAP（分泌碱性磷酸酶），萤光素酶，β-半乳糖苷酶和GFP（绿色 荧光蛋白）与选自NPT（新霉素磷酸转移酶），CAT（氯霉素乙酰转移酶）和β-半乳糖苷酶，lacZ的选择标记表达基因; 将其转染至皮肤细胞系; 并通过荧光测定法定量测定来自上述转染的皮肤细胞系（KCLRF-BP-00035）的NF-κB的表达。