Expression of Human eNOS in Cardiac and Endothelial Cells

: In this chapter we provide a detailed technique-focused protocol for expression of human endothelial nitric oxide synthase (eNOS) gene delivered by replication-deficient adenovirus. It includes construction of recombinant plasmid with human eNOS gene full-length cDNA (heNOS), packaging of adenovirus with heNOS (Adv-heNOS) in 293 cells, confirmation of Adv-heNOS, amplification of high-titer stockAdv-heNOS, determination of the Adv-heNOS titer, expression of Adv-heNOS in the human abdominal aorta endothelial (HAAE1) cell line and rat heart myoblast cell line (H9C2), and measurement of human eNOS activity and NO production.