Figure 2.

Construction and characterization of copGFP-expressing adenoviral vector using the OSCA system

(A) Construction of AdOS-copGFP using the OSCA system. The copGFP coding sequence was PCR amplified with MOS1- and MOS2-anchored primers (a), followed by Gibson Assembly (b). (B) Bacterial colonies post the Gibson Assembly reaction. (C) Identification of pAdOS-copGFP using PCR screening of bacterial colonies. Randomly picked up 16 colonies were PCR amplified with copGFP specific primers, and all but one (#11) were positive for copGFP. (D) Validation of adenoviral recombinant pAdOS-copGFP clones. The representative three clones, along with the control adenoviral backbone vector pAdEasy1, were digested with Hind III (a), Kpn I (b), Bam HI (c), and Sph I (d). The digested plasmid DNA was resolved in 1% agarose gels.