Emergence of large plaque-producing clones of human immunodeficiency virus (HIV) in vitro.
Large plaque-inducing clones were obtained from small plaque-inducing parental clones of human immunodeficiency virus (HIV) by the plaque-cloning method. The cloned HIVs that formed large and small plaques were studied as follows: 1) infectivity was determined by the ratio of plaque-forming units (PFU) to reverse transcriptase (RT) activity; 2) viral growth was assessed by the amount (RT activity) of virus after infection; and 3) HIV long terminal repeat (LTR)-linked gene expression of the viruses was measured by chloramphenicol acetyltransferase (CAT) assay using persistently infected MOLT-4 cells. Results showed that clones producing large plaques showed similar or slightly lower infectivity but higher virus production, faster viral growth, and higher gene expression activity than clones producing small plaques. These analyses revealed that clones producing large plaques could replicate more rapidly than those producing small plaques. Restriction enzyme map analysis of these cloned viruses showed that they were also genetically different. These results suggest that the changes in the biological features observed here might be due to mutation during the cloning procedure.
Masuda
T
T
Institute for Virus Research, Kyoto University, Japan.
Kannagi
M
M
Nakamura
M
M
Ohtani
K
K
Hinuma
Y
Y
Harada
S
S
D004279:DNA, Viral / Q000032:analysis; D006678:HIV / Q000235:genetics / Q000254:growth & development*; D009154:Mutation; D010948:Viral Plaque Assay; D014779:Virus Replication
D004279:DNA, Viral