Skip to main content
. Author manuscript; available in PMC: 2010 Feb 1.
Published in final edited form as: Expert Rev Vaccines. 2009 Apr;8(4):469–481. doi: 10.1586/erv.09.1

Figure 2. The AdHigh system.

Figure 2

The pAdHigh shuttle plasmid is a hybrid between pShuttle-CMV [79] and pAdApt [35], containing Ad5 sequences flanking an expression cassette. After cloning a transgene into the cassette, homologous recombination is allowed to occur between the shuttle plasmid pAdHigh and the Ad5 backbone plasmid pAdEasy-1 in Escherichia coli BJ5183 cells. RCA-free Ad5 vectors encoding a transgene can be generated by transfecting PER.C6 cells with the recombinant plasmid.

Ad5: Adenovirus serotype 5; Amp: Ampicillin-resistance gene; CMV: Cytomelagovirus; Kan: Kanamycin-resistance gene; Left arm: Ad5 nucleotides 34931–35935; LITR: Ad5 nucleotides 1–454; MCS: Multiple cloning site; polyA+: Simian virus 40 polyadenylation signal; Ori: PBR322 origin of replication; Promoter: CMV promoter; RCA: Replication-competent adenovirus; Right arm: Ad5 nucleotides 3511–5790 including a functional pIX promoter [81].

This schema was a modification of that for generating AdEasy-derived Ad5 vectors in 293 cells [79].