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Gene Synthesis> | Human DYK tagged O-GlcNAc transferase (OGT, OHu54262) and pig Pgc1α-myc (OSe00030D; p.C430S) clones were obtained from GenScript. Pgc1α was subcloned into pEGFP-N1 using BamHI and XhoI restriction enzymes. Site-directed mutagenesis was performed using Pfu Ultra II HS DNA polymerase (Agilent) and DpnI (Fermentas). The following primers were used to create p.430C variant in pig Pgc1α (PGC1aSer430Cys-F 5’-ccacagactcagaccagtgctacctgaccgagac gtcggag-3’ and PGC1aSer430Cys-R 5’-ctccgacgtctcggtcaggtagcactggtctgagtctgtgg-3’). The stop codon was eliminated to overexpress Pgc1α-myc-GFP using PGC1aEGFP-F 5-catctcagaagagga tctgttggatccaccggtcgccacc-3 and PGC1aEGFP-R 5-ggtggcgaccggtggatccaacagatcctcttctgagatg-3. | Get A Quote |
PGC1α is a coactivator of many transcription factors and cytosolic phosphoenolpyruvate carboxykinase (PCK1) is a key enzyme for gluconeogenesis. PGC1α interacts with the transcription factor PPARγ to stimulate PCK1 expression and thus de novo glucose synthesis. These proteins are not only important for central energy metabolism but also for supplying intermediates for other metabolic pathways, including lipidogenesis and protein synthesis and might therefore be important factors in the ethiopathogenesis of metabolic disorders like diabetes but also in other pathologies like cancer. Since polymorphisms in these proteins have been related to some phenotypic traits in animals like pigs and PGC1α G482S polymorp... More