Conditional replication of a recombinant adenovirus studied using neomycin as a selective marker.
An E1B-defective adenovirus, named r2/Ad carrying the neo expression cassette, was constructed by homologous recombination. The construction, selection (using neomycin as a selective marker), and propagation of the recombinant virus was performed in human embryonic kidney 293 cells (HEK 293). An in vitro study demonstrated that this recombinant virus has the ability to replicate in and lyse some p53-deficient human tumor cells such as human glioma tumor cells (U251) and human bladder cells (EJ), but not in some cells with functional p53, such as human adenocarcinoma cells (A549) and human fibroblast cells (MRC-5). Also, based on the cytopathic effect (CPE), it was demonstrated, under identical conditions, that the U251 cells were more sensitive to r2/Ad replication than the EJ cells. In this paper, we report that r2/Ad could be very useful in studying the in vitro selective replication of E1B-defective adenovirus and has great potential in cancer gene therapy.
Xue
Feng
F
Institute of Virology, Wuhan University, Wuhan 430072, P. R. China.
Qi
Yi-Peng
YP
Joshua
Mallam Nock
MN
Lan
Ping
P
Dong
Chang-Yuan
CY
D000256:Adenoviridae / Q000187:drug effects / Q000235:genetics*; D017501:Adenovirus E1B Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D004352:Drug Resistance, Microbial / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005819:Genetic Markers; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D019868:Kanamycin Kinase / Q000235:genetics* / Q000378:metabolism; D009355:Neomycin / Q000494:pharmacology*; D011995:Recombination, Genetic; D016159:Tumor Suppressor Protein p53 / Q000235:genetics; D014779:Virus Replication / Q000187:drug effects / Q000235:genetics*
D017501:Adenovirus E1B Proteins; D005819:Genetic Markers; D016159:Tumor Suppressor Protein p53; D019868:Kanamycin Kinase; D009355:Neomycin