Molecular Biology and Biochemistry Core

核心C是分子生物学和生物化学的核心。这个核心负责协助每个 在分离的肾节段的显微解剖、基因分型、基因沉默和 定量检测mRNA和蛋白质的表达。基因分型将用实时荧光聚合酶链式反应进行 在小鼠上使用ABI PRISM 7700 HT序列检测系统，其中以下基因 已经被破坏：NADPH氧化酶的p22Phox亚单位；细胞外超氧化物歧化酶 超氧化物歧化酶(EC-SOD)、细胞内超氧化物歧化酶(IC-SOD)、锰超氧化物歧化酶(Mn-SOD) 超氧化物歧化酶)、血管紧张素1a型受体(AT-1AR)、多巴胺5型受体(D5R)、成纤维细胞生长 第二型和第七型成纤维细胞生长因子和成纤维细胞生长因子结合蛋白(BP1)被敲除 转基因。核心C还将通过siRNA帮助沉默以下基因：BP1、FGF2和FGF2- 7.此外，Core C将通过实时荧光聚合酶链式反应、微阵列 原位杂交和蛋白印迹、免疫组织化学检测蛋白表达。 放射配基结合和放射自显影，以评估siRNA的有效性和特异性。这 CORE还将负责向调查人员通报最新的分子技术 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.