背景介绍 Phosphodiesterases (PDEs) regulate the intracellular levels of cAMP and cGMP by hydrolyzing cAMP and cGMP to their inactive 5' monophosphates. These cyclic nucleotides play an important role as second messengers in diverse physiological functions. PDE7 is a cAMP-specific enzyme and two PDE genes (PDE7A and PDE7B) have been identified. PDE7 is widely expressed in various tissues, with PDE7A found primarily in skeletal muscle, T lymphocytes, and pancreas, while high levels of PDE7B are detected in brain, heart, and liver. Inhibition of PDE7 activity by its inhibitors leads to elevated intracellular level of cAMP. 产品介绍 Recombinant HEK293 cell line expressing rat PDE7A (phosphodiesterase 7A, accession number NM_031080).
N-terminal FLAG-tagged rat PDE7A has been stably expressed in a human embryonic kidney (HEK293) cell line and its expression was confirmed by Western blotting. The regulation of intracellular level of cAMP by rat PDE7A in rat PDE7A stably-expressed HEK293 cells was characterized by a cell-based reporter assay using pCRE-luc reporter vector. pCRE-luc contains a luciferase gene that is under the control of the cAMP response element (CRE). When cells transiently transfected with pCRE-luc reporter were activated by forskolin, the level of cAMP was upregulated in parental HEK293 cells inducing the expression of luciferase reporter, whereas rat PDE7A-HEK293 cells showed reduction in the level of cAMP, resulting in lowered levels of luciferase expression. Inhibition of PDE7A activity by BRL 50481, a PDE7A inhibitor, restored the cAMP level, resulting in higher luciferase activity.