TABLE 1. PCR analysis of tissues from baboons injected with ADV/RSV-tk. ND = not 
done. 
ORGAN 
Mod. dose 
+ GCV; _ 
Mod. dose 
+ GCV; _ 
High dose 
+ GCV; _ 
High dose 
+ GCV; _ 
High dose 
- GCV; _ 
High dose 
- GCV; _ 
Injection site 
- 
- 
+ 
+ 
+ 
- 
CNS distal to site 
- 
- 
+ 
- 
- 
- 
Lung 
ND 
- 
ND 
- 
- 
- 
Kidney 
- 
■ 
- 
- 
- 
ND 
Ovary/Testis 
★ 
- 
- 
- 
- 
* Removed before study. 
FIGURES 
Construction of Recombinant Adenovirus Vector 
Containing the HSV-tk Gene 
p J M 17 
X 
Fig. 1 Cloning strategy for 
ADV/RSV-tk. The 2.8 kbp Bgl II to 
BamH I fragment containing the 
HSV-tk gene and poly-A tail was 
inserted into the BamH I site of the 
plasmid pADL.I/RSV previously 
constructed in Dr. Woo's laboratory 
by Dr. B. Fang to generate the 
plasmid pADL.I/RSV-tk. To produce 
the recombinant adenovirus, 
pADL.I/RSV-tk and pJM17, a 
plasmid containing the adenovirus 
genome modified so that it is too 
large to be packaged (supplied by 
Dr. Frank Graham of McMaster 
University), were co-transfected into 
the 293 transformed human kidney 
cell line by the calcium phosphate 
method. Recombinant adenovirus was isolated from a single plaque, expanded in the 293 
cell line and purified by cesium gradient ultracentrifugation (Graham and Prevec, 1991). 
pADL.I/ RSV-tk 
HSV-tk 
- ) HSV-tk 
Co-transfection into 
293 cells (E1+) 
AD V/ RSV-tk 
Recombinant DNA Research, Volume 20 
[731] 
