M.J. Welsh and A.E. Smith, RAC Application 
Clal(i) 
Aatll 
Ed I 361(8 I 6) 
Bell 36 1(904) 
Spel(9 1 5) 
PBR322 
BamHI 
(7927) 
BstBI 
BamHI 
(12427) 
BstBI 
Eel l 361/SnaBI 
Ad2-3498 
A fill 
Figure 3. Construction of the Ad2/CFTR-1 vector 
CFTR eDNA within this plasmid has been completely sequenced. The Spel/Ecll36I 
restriction fragment contains 47 bp of 5' sequence derived from synthetic linkers and the 
multiple cloning site of the vector. 
The CFTR cDNA was inserted between the Nhel and SnaB 1 restriction sites of the 
adenovirus gene transfer vector pBR-Ad2-7. pBR-Ad2-7 is a pBR322 based plasmid 
containing an approximately 7kb insert derived from the 5' 10680 bp of Ad2 inserted 
between die Clal and BamHI sites of PBR322. From this Ad2 fragment, we have deleted 
sequences corresponding to Ad2 nucleotides 546-3497 and replaced them with a 12 bp 
multiple cloning site containing an Nhel site, an Mlul site, and a SnaBl site. The construct 
also contains the 5' inverted terminal repeat and viral packaging signals, the El a enhancer 
and promoter, the Elb 3' intron and the 3' untranslated region and polyadenylation sites. 
The resulting plasmid was called pBR-Ad2-7/CFTR. Its use to assemble virus is described 
in Blb(l)b. 
Recombinant DNA Research, Volume 16 
[875] 
