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Journal: یاخته
Year:2008 | Volume: | Issue:
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Title

PROFILE ANALYSIS OF PEROXISOMAL MARKER GENES EXPRESSION IN COMPARISION WITH OCT4 AND NANOG DURING NEURAL DIFFERENTIATION OF P19 CELLS

Pages

 Start Page 87 | End Page 87

Abstract

 Objective: PEROXISOMEs are ubiquitous organelles in eukaryotes that participate in the metabolism of fatty acids and other metabolites; they also play important roles in neurogenesis and biogenesis. In order to investigate PEROXISOME biogenesis profile during neurogenesis, we have performed to set a semi quantitative analysis for two peroxisomal genes expression (Catalase and PEX3) comparing with STEM CELL marker genes (Oct4 and Nanog) and a house keeping gene (b -actin) in P19 cells. In this project we have used P19 cells which are suitable progenitor cells for the study of neurogenesis. Moreover two different peroxisomal genes were selected to chase both kinetics of peroxisomal matrix protein synthesis (Catalase) and peroxisomal membrane protein biogenesis (Pex3p). Total RNA extracted from P19 cells and was used for cDNA synthesis at the next step.Materials and Methods: Using specific primers a part of OCT4, Nanog, b-actin, PEX3 and Catalase cDNAs were amplified by RT-PCR for quantitative analysis.Results: Various parameters were changed to optimize the PCR profiles for each gene for further assessments. Here we describe the conditions which we used in detail.Conclusion: PEX3 band was observed sharply and amplified at PCR conditions of annealing temperature of 55.1oc, we got b -actin band at 51oc to 55oc degree for annealing. For PCR of Catalase annealing temperature of 52.7°c was used, Nanog band was at 66oc and OCT4 band was observed at 51oc to 57oc.

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