Generation of ACN53 
xsar e srt2 4 * 3 s 
NmmaCUVC< 
L— dr 
Isolate 4.6 Kb 
Eel 136 11/NgoM I 
Fragment 
5000 10000 15000 20000 
»000 35000 
NgoMl 
Oral 
Oral 
NgoWi 
pNgoMI 
NgoMl 
[NgoMl 
fNgoMI 
rwgoMi 
ktf.^ggwB^ 
-NgoMl 
-NgoMl 
r Oral [Oral 
[NgoMl 
NgoMl 
Isolate 6437 bp Dra 1/NgoM I 
Fragment (Ad2 4021-10457) 
["NgoMl 
[Oral 
NgoMl 
AdZ 
«n m i+m7*Ata 
Mix 
Ligate 
Transform E. coli DH5a 
t 
AAd 357-4020 
EcoR I digest 
Cla 
AE3 
Ad5/dl327 
I 
Cla I digest 
purify large fragment 
Mix and transfect 293 cells 
In vivo recombination 
5000 
10000 
15000 
20000 
25000 
30000 
I I AE3 
Delete Ad5 El a, Elb, pIX 
Inserted CMV/TPl7p53 cDNA 
ACN53 
[ 694 ] 
Recombinant DNA Research, Volume 20 
