CORE--IMMUNOLOGY AND MOLECULAR BIOLOGY FACILITY

核心——免疫学和分子生物学设施

• 批准号: 6105874
• 负责人: NORMAN A GRANHOLM
• 金额: $ 16.43万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-03-01 至 2000-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6105874
• 关键词: HMG coA reductases; antibody; biomedical facility; genetic screening; genotype; immunocytochemistry; immunology; in situ hybridization; low density lipoprotein receptor; messenger RNA; microorganism culture; molecular biology; polymerase chain reaction

This Core Laboratory will provide support in five areas: (1) to produce, maintain, and characterize antibodies, (2) to analyze genomic DNA by polymerase chain reaction, (3) to analyze and to quantitate cellular mRNA, (4) to provide histopathology, immunohistochemistry, and in situ hybridization support, and (5) to maintain bacterial culture and plasmid stocks. (1st Core Area). Western blots are used to characterize antibody specificity and cross reactivity to pancreatic lipases, cholesterol esterase, phospholipase A/2, and to lipid binding proteins though to be involved in lipid uptake by the epithelial cells of the small intestine. These antibodies are used in Western blots, enzyme linked immunosorbent assays, and in immunohistochemical studies to measure changes in protein amount and location that occur in tissues and fluids in response to dietary manipulations, gene knock out, and ectopic expression of pancreatic lipase. (2nd Core Area). The progeny of targeted gene knock out mice are genotyped by polymerase chain reaction. Genotyping of human DNA is performed to assess apoE alleles and apoA-IV alleles in a population of potential study patients. (3rd Core Area). Cellular mRNA for human low density lipoprotein receptor and for hydroxymethylglutaryl CoA reductase is characterized by reverse transcription polymerase chain reaction. The changes in mRNA responses to dietary manipulations are characterized. Additionally, in mice the impact of knock out or of ectopic expression of pancreatic lipase on dietary regulated expression of pancreatic lipases and cholesterol esterase is characterized. (4th Core Area). We will provide support services for histopathological studies, immunohistochemical studies, and in situ hybridization studies of tissues. (5th Core Area). We will maintain and distribute bacterial cultures and plasmid stocks in this Core.

该核心实验室将在五个方面提供支持：（1）生产， 维护和表征抗体，（2）通过以下方法分析基因组DNA： 聚合酶链反应，（3）分析和定量细胞 mRNA，（4）提供组织病理学、免疫组织化学和原位 杂交支持物，和（5）维持细菌培养物和质粒 名的股票明细金额 (1st核心区）。蛋白质印迹用于表征抗体 对胰脂肪酶、胆固醇 酯酶，磷脂酶A/2，和脂质结合蛋白，虽然要 参与小肠上皮细胞的脂质摄取。 这些抗体用于蛋白质印迹、酶联免疫吸附 在免疫组化研究中， 在组织和体液中发生的数量和位置 饮食控制、基因敲除和异位表达 胰脂肪酶 (2nd核心区）。靶向基因敲除小鼠的后代是 通过聚合酶链反应进行基因分型。人类DNA的基因分型是 进行评估apoE等位基因和apoA-IV等位基因在人群中， 潜在的研究患者。 (3rd核心区）。人低密度脂蛋白的细胞mRNA 受体和羟甲基戊二酰辅酶A还原酶的特征在于： 逆转录聚合酶链反应mRNA的变化 表征了对饮食操作的反应。此外，在 小鼠胰腺癌基因敲除或异位表达的影响 脂肪酶对膳食调节的胰脂肪酶表达的影响， 胆固醇酯酶的特征。 (4th核心区）。我们将提供组织病理学支持服务， 免疫组化研究和原位杂交研究 组织。 (5th核心区）。我们将保存和分发细菌培养物， 在这个核心中的质粒储备。