Molecular cloning of novel recombinant plasmids for exogenous expression of PKCε in single living cells

• Main Author: Su, Dacia Luan Yu.
• Other Authors: Chen Peng
• Format: Final Year Project
• Language: English
• Published: 2009
• Subjects: DRNTU::Engineering::Chemical engineering::Biotechnology
• Online Access: http://hdl.handle.net/10356/16545
• Institution: Nanyang Technological University

In this study, overexpression was done on the recombinant plasmids of both wild-type PKCε (wt-PKCε) and dominant negative PKCε (dn-PKCε) in PC 12 cells with reporter genes, Enhanced Green Fluorescent Protein at C1 position (pEGFP-C1), pIRES2-EGFP and pIRES2-DsRed. Both fusion and concurrent gene expression were carried out and successful recombinant cells expressing wt-PKCε and dn-PKCε genes in mammalian cell cultures, were identified via screening for fluorescence in microscopy one day after transfection. As compared to employing Western blotting that is usually used on dead cells, this study allows for a proper report in living cells.