Figure 2.4-C. Construction of AvlCFl (a recombinant adenovirus expressing 
human CFTR used for transferring the CFTR cDNA) by homologous recombination 
of the E3 deletion mutant Ad-dl327 and plasmid pS2CFTR. The majority of the 
E3 region has been deleted in Ad-dl327 ; there is also a minor 2 bp deletion 
of the VA-I promoter (region of 29 m.u. ; not shown). The site of the Ela 
region is indicated by the horizontal arrow. Within the site is a conve- 
nient Clal site. Ad-dl327 is digested with Clal to yield the "Clal large 
fragment" . The shuttle plasmid pS2CFTR is cleaved with Notl and Kpnl and 
co- transfected with the Clal large fragment into 293 cells where homologous 
recombination occurs to yield the infectious deficient recombinant adenovi- 
rus AvlCFl. At the bottom is shown AvlCFl (total length 101 m.u.) with an 
enlarged view of the left end. These elements are identical to those in 
pS2CFTR, see Figure 2.4-B. 
Map units 
0 10 20 30 40 50 60 70 80 90 100 
1 1 1 1 1 1 1 1 1 1 1 
Ad 5 
9.24 17.34 
Human 
CFTR Ad 5 
cDNA 
Transfect into 
293 cells 
AvlCFl 
Map units 
0 10 20 30 40 50 60 70 80 90 100 
1 1 1 1 1 1 1 1 1 1 n 
101 
ITR 
Origin of 
replication 
Ela promoter/enhancer 
and encapsidation 
signal (d») 
i i 
500 bp 
[ 666 ] 
Recombinant DNA Research, Volume 16 
