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L-type Ca channels (LTCCs) are the main mediators of Ca influx. LTCCs are multi-subunit complexes, of which CaV1.2 subunit forms the channel pore for Ca movement. Ca influx through LTCC CaV1.2 is a proximal signal for pathological cardiomyocyte hypertrophy. However, very little is known regarding the molecular mechanism of LTCC transcription regulation. Myocardin, a co-activator of serum response factor (SRF) that potently transactivates CArG box–containing cardiac and smooth muscle target genes, can stimulate neonatal rat cardiomyocyte hypertrophy. In the present study, the gene fragment of rat LTCC CaV1.2 promoter containing five CArG boxes or no CArG box was amplified by PCR and cloned into empty vector pGL3-Basic to construct luciferase reporter plasmids, then luciferase activity assay was performed in HEK293T cells. The results showed that the luciferase reporter plasmids were constructed successfully, Myocardin potently activated CArG boxcontaining LTCC CaV1.2 reporter, by contrast, Myocardin had a weaker transcriptional activation on LTCC CaV1.2 reporter containing no CArG box. The results suggest that Myocardin may play a critical role in regulating the expression of LTCC CaV1.2 in cardiomyocyte hypertrophy. Keywords-CaV1.2; CArG box; promoter; luciferase activity assay

Construction and Functional Analysis of Luciferase Reporter Plasmid Containing CaV1.2 Gene Promoter