Molecular Biology and Biochemistry Core

核心C是分子生物学和生物化学核心。这个核心负责协助每个 在离体肾段的显微切割，基因分型，基因沉默， mRNA和蛋白质表达的定量。将采用实时PCR进行基因分型 在小鼠上使用ABI PRISM 7700 HT序列检测系统，其中 已被破坏：NADPH氧化酶的p22 phox亚基;细胞外超氧化物歧化酶 （EC-SOD）、胞内超氧化物歧化酶（IC-SOD）、锰超氧化物歧化酶（Mn- SOD）、血管紧张素1a型受体（AT-1aR）、多巴胺5型受体（D5 R）、成纤维细胞生长 因子2（FGF-2）和因子7（FGF-7）以及FGF结合蛋白（BP 1）敲除， 转基因。核心C还将通过siRNA帮助沉默以下基因：BP 1、FGF-2和FGF-10。 7.此外，核心C将有助于通过实时PCR、微阵列技术和基因芯片技术确定mRNA表达。 原位杂交和Western印迹，免疫组织化学， 放射性配体结合和放射自显影以评估siRNA功效和特异性。这 核心还将负责使研究人员了解最新的分子技术 适用于每个子项目。

Core C is the Molecular Biology and Biochemistry Core. This core is responsible for assisting each of the subprojects in microdissection of isolated kidney segments, genotyping, gene silencing, and in quantitation of mRNA and protein expression. Genotyping will be performed with real-time PCR using an ABI PRISM 7700 HT sequence detection system on mice in which the following genes have been disrupted: the p22phox subunit of NADPH oxidase; extracellular superoxide dismutase (EC-SOD), intracellular superoxide dismutase (IC-SOD), manganese superoxide dismutase (Mn- SOD), angiotensin type 1a receptor (AT-1aR), dopamine type 5 receptor (D5R), fibroblast growth factor type 2 (FGF-2) and type 7 (FGF-7) and the FGF binding protein (BP1) knockout and transgene. Core C will also assist in silencing the following genes by siRNA: BP1, FGF-2 and FGF- 7. In addition, Core C will assist in determining mRNA expression by real-time PCR, microarray and in situ hybridization and in assaying protein expression by Western blot, immunohistochemistry, radioligand binding and autoradiography in order to assess siRNA efficacy and specificity. This core will also be responsible for keeping investigators informed of the latest molecular techniques applicable for each of the subprojects.