MAMMALIAN CELLS EXPRESSING FMO

表达 FMO 的哺乳动物细胞

• 批准号: 2545785
• 负责人: CHARLES L CRESPI
• 金额: $ 19.8万
• 依托单位: GENTEST CORPORATION
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-30 至 1999-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2545785
• 关键词: animal tissue; complementary DNA; cytochrome P450; drug metabolism; enzyme activity; flavoproteins; gene expression; human genetic material tag; lymphoblast; method development; mutagen testing; neoplastic transformation; oxygenases; tissue /cell culture; toxicology; transfection /expression vector; western blottings

Rabbit FM01 and FM02 and human FM03 cDNAs will be expressed in AHH-1TK+/- human lymphoblastoid cells, AS52 Chinese hamster cells and C3H 10T1/2 mouse cells. FMO expression will be monitored by regio- and stereoselective/enzyme assays and immunoblotting. The level and stability of FMO expression will be monitored in the cDNA expressing cell lines. The differences in xenobiotic metabolism by allelic variants of human FM03 will be examined. The FMO-expressing human lymphoblastoid cell lines will compliment an existing panel of cell lines expressing cytochrome P450 enzymes. AS52 cells will allow the facile analysis of mutational spectrum for promutagens activated by FMO. The C3H 10T1/2 cell lines will allow extension of in vitro toxicology studies to the malignant transformation endpoint. PROPOSED COMMERCIAL APPLICATION: Mammalian cells expressing flavin- containing monoxygenases will have commercial applications for the in vitro analysis of xenobiotic metabolism and toxicity. This new system complements an existing system containing cytochrome P450 enzymes.

兔FM 01和FM 02以及人FM 03 cDNA将在AHH-1 TK +/- 人淋巴母细胞、AS 52中国仓鼠细胞和C3 H 10 T1/2小鼠 细胞 FMO表达将通过区域和 立体选择性/酶测定和免疫印迹。 水平和稳定性 将在cDNA表达细胞系中监测FMO表达。 的 人FM 03的等位基因变体的异生物质代谢的差异将 接受检查。 表达FMO的人淋巴母细胞样细胞系将 补充了现有的一组表达细胞色素P450的细胞系 内切酶 AS 52细胞将允许突变谱的简易分析 FMO激活的前诱变剂 C3 H 10 T1/2细胞系将允许 将体外毒理学研究扩展至恶性转化 终点。 建议的商业应用：表达黄素的哺乳动物细胞- 含有单加氧酶的化合物将具有体外 异生物质代谢和毒性分析。 这个新系统 补充了含有细胞色素P450酶的现有系统。