This kit supplies reagents sufficient to make 5 recombinant adenovrial vectors. It is intended for those who wish to select a promoter as well as the gene to be expressed. A polyadenylation signal is also required. All 3 sequences are inserted into the multiple cloning site of pLEP and the plasmid is then cut with a restriction enzyme (PI-Psp I) and ligated to the remainder of the adenoviral genome, which is provided in a pREP plasmid. Two versions of the kit are available, one with pREP and the other with pREP-E3. Vectors made with pREP can accommodate inserts up to 5 kb, whereas those made with pREP-E3 can accommodate even longer inserts (up to 8 kb). The additional capacity is due to deletion of the adenoviral E3 genes. The E3 genes reduce host immune responses to viral infection, but they are not essential for viral propagation. We recommend using pREP whenever possible. The pREP plasmid has been linearized and is ready to be ligated to pLEP. For those who desire high level gene expression in all tissues, we recommend using the pLEP-CMV vector instead of pLEP (see Kit #K1100, SpeAd^TM -CMV kit). The manual that accompanies the kit provides detailed instructions for making adenoviral vectors using the SpeAd^TM method.

Kit includes: pLEP (20 ¦Ìg)

pREP7 (or 8), linearized (10 ¦Ìg)

positive control plasmid to express b-galactosidase (10 ¦Ìg)

lambda phage packaging extract (5 x 50 ¦Ìl)

phage dilution buffer (3 ml)

ready-to-use bacterial host (E coli DK-1 cells, 1 ml)

sequencing primers (2, 100 ng each)

SpeAd^TM Manual