pET17b is a prokaryotic expression vector (Novagen). It is used to obtain restriction enzyme sites XbaI and NotI, and to make it easy to insert the target gene into the pShuttle vector for packaging Adv-heNOS. The pShuttle expression system (BD Biosciences Clontech) allows the target gene into a mammalian expression cassette, which consists of the human CMV immediate-early promoter/enhancer(PCMV IE), a multiple cloning site (MCS), and the bovine growth hormone polyadenylation signal (BGH polyA). The entire cassette is flanked by unique I-CeuI and Pl-SceI restriction enzyme sites so that it can be excised and ligated to Adeno-X viral DNA. The vector backbone also possesses the pUC origin (pUC ori) and a kanamycin resistance gene (Kanr) for propagation and selection in E. coli.
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