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Emergence of large plaque‐producing clones of human immunodeficiency virus (HIV) in vitro
Author(s) -
Masuda Takao,
Kannagi Mari,
Nakamura Masataka,
Ohtani Kiyoshi,
Hinuma Yorio,
Harada Shinji
Publication year - 1989
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.1890270220
Subject(s) - biology , virology , chloramphenicol acetyltransferase , infectivity , virus , virus quantification , cloning (programming) , reverse transcriptase , plaque forming unit , long terminal repeat , microbiology and biotechnology , gene , restriction enzyme , viral replication , in vitro , gene expression , polymerase chain reaction , genetics , reporter gene , computer science , programming language
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.