Shuttle plasmid for the construction of replication-deficient, RCA-free adenovirus vectors using the AdenoQuick2.0 system (E1 deletion, complete packaging signal)
- Derivative of pAd1127-02
- Complete packaging signal including the 7 "A"-repeats
- Insertion of heterologous sequences in place of E1 region of the Ad5 genome
- Easy manipulation of pIX region
pAd1127-06 is a plasmid designed for inserting expression cassettes in place of the E1 region of the Ad5 genome and to manipulate the pIX promoter and coding region. It contains PacI and SwaI sites flanking the first 440 base pairs from the Ad5 genome (including the left ITR and packaging signal), a multiple cloning site, and the pIX coding region. Expression cassettes inserted into the multiple cloning site should contain a promoter, coding sequence and a polyA signal. The sequences encompassing the kanamycin-resistance gene, the λ cos site, the adenovirus 0-1.3 map units, the multiple cloning site and the pIX coding sequence are flanked by two SfiI restriction sites. These sites generate non-symmetrical sticky ends suitable for directional cloning with the other AdenoQuick2.0 plasmids (pAd1128, pAd1129, pAd1130, and their derivatives).
pAd1127-06 is a derivative of pAd1127-02, in which the packaging signal has been extended from psn 350 to psn 440 (in the Ad5 genome), to include all 7 packaging "A" repeats (I, II, III, IV, V, VI, and VII). The complete packaging signal region might confer a growth advantage to the virus, according to Youil et al (Human Gene Therapy 14: 1017-1034). Because of the size of the E1 deletion (440-3510), the vectors generated from pAd1127-06 have minimal or no homology with the Ad5 sequences inserted in the chromosome of the helper cells such as PER-C6, thereby minimizing the probability of RCA generation.
The adenovirus sequences contained in pAd1127-06 have been verified by sequencing.
|origin of replication||1698-1110||pUC19|
|Vector Map||Sequence||Product Information Sheet|
|Vector Map||pAd1127-06.txt (3.08 KB)||Product_Information_pAd1127-06.pdf (227.77 KB)|