Shuttle plasmid for the construction of adenovirus vectors using the AdenoQuick2.0 system (2.7 kb E3 deletion and Ad5/3 fiber)
- 2.7 kb BglII fragment deleted from E3 region and replaced with multiple cloning site
- Dicistronic (bipartite) adenovirus vectors
- "Armed" conditionally-replicative (oncolytic) adenovirus vectors (CrAds)
- Easy manipulation of fiber coding sequence
pAd1129-27 is a shuttle plasmid designed for constructing adenovirus vectors characterized by a 2.7 kb deletion in the E3 region, and a hybrid Ad5/3 fiber. A 2.7 kb BglII fragment including E3 6.7K, gp19K membrane protein, the adenovirus “death” protein ADP, RID-a, RID-ß, and14.7K was deleted, and replaced with a multiple cloning site. The E3 12.5K ORF is truncated. The U exon is intact. The Ad5/3 hybrid fiber is made of the N-terminal tail and shaft of Ad5 fused to the knob of Ad3.
pAd1129-27 can be used to construct replication-deficient or oncolytic adenovirus vectors expressing large transgenes (inserted into the E3 region itself or elsewhere), or multiple expression cassettes (for instance two independent expression cassettes, one in the E1 region, and the other in the E3 region). Expression cassettes inserted into the E3 region should contain a promoter and poly(A) signal, but no intron nor splice site. The adenovirus sequences present in pAd1129-27 are flanked by two SfiI sites, which generate non-symmetrical sticky ends suitable for directional cloning with the other AdenoQuick2.0 plasmids (pAd1127, pAd1128, pAd1130, and their derivatives).
|Sequence + Annotations
|pAd1129-27.txt (4.19 KB)
|pAd1129-27.gb (9.52 KB)