Molecular Biology and Biochemistry Core

$193,704P01FY2008HLNIH

Georgetown University, Washington DC

Investigators

Linked publications & trials

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Abstract

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

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