CORE--IMMUNOLOGY AND MOLECULAR BIOLOGY FACILITY

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) 通过以下方式分析基因组 DNA： 聚合酶链式反应，(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，以及 (5) 维持细菌培养物和质粒 股票。 （第一核心区）。蛋白质印迹用于表征抗体 对胰脂肪酶、胆固醇的特异性和交叉反应性 酯酶、磷脂酶 A/2 和脂质结合蛋白 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 测定，以及在免疫组织化学研究中测量蛋白质的变化 组织和体液中发生的响应的数量和位置 饮食控制、基因敲除和异位表达 胰脂肪酶。 （第二核心区）。靶向基因敲除小鼠的后代是 通过聚合酶链式反应进行基因分型。人类 DNA 的基因分型是 评估人群中的 apoE 等位基因和 apoA-IV 等位基因 潜在的研究患者。 （第三核心区）。人低密度脂蛋白的细胞 mRNA 羟甲基戊二酰辅酶A还原酶受体的特征在于 逆转录聚合酶链反应。 mRNA的变化 对饮食控制的反应是有特征的。另外，在 小鼠胰腺基因敲除或异位表达的影响 脂肪酶对胰腺脂肪酶饮食调节表达的影响 胆固醇酯酶被表征。 （第四核心区）。我们将为组织病理学提供支持服务 研究、免疫组织化学研究和原位杂交研究 组织。 （第五核心区）。我们将维护和分发细菌培养物， 该核心中的质粒库存。

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.