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， 维护和鉴定抗体，(2)通过以下方式分析基因组DNA 聚合酶链式反应，(3)细胞分析和定量 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)保持细菌培养和质粒 股票。 (第一核心区)。用免疫印迹法鉴定抗体 对胰腺脂肪酶、胆固醇的特异性和交叉反应性 酯酶、磷脂酶A/2和脂结合蛋白 与小肠上皮细胞的脂肪摄取有关。 这些抗体用于免疫印迹、酶联免疫吸附试验 检测，以及在免疫组织化学研究中测量蛋白质的变化 在组织和体液中出现的数量和位置 饮食操作、基因敲除和异位表达 胰腺脂肪酶。 (第二核心区)。靶向基因敲除小鼠的后代是 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 潜在的研究对象。 (第三核心区)。人低密度脂蛋白的细胞基因表达 受体和羟甲基戊二酰辅酶A还原酶的特征是 逆转录聚合酶链式反应。核糖核酸的变化 对饮食控制的反应进行了表征。此外，在 基因敲除或异位表达对小鼠胰腺的影响 脂肪酶对饮食调节胰腺脂肪酶基因表达的影响 对胆固醇酯酶进行了表征。 (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 纸巾。 (第五核心区域)。我们将维护和分发细菌培养物和 在这个核心中储存了质粒库。

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.