Development of RD-tat cell lines: use in HIV recombination studies.
The transactivator (tat) gene of human immunodeficiency virus (HIV) plays an essential role in the replication cycle of HIV. Previous studies have evaluated the extent and mechanistic aspects of tat-mediated transactivation using lymphoid and adherent non-lymphoid cells. We have exploited the transactivation property of the tat gene to achieve high levels of hybrid HIV resulting from recombination between HIV DNAs. For this purpose, we have generated stably transformed human rhabdomyosarcoma (RD) cell lines expressing tat gene product of HIV-1. Functional analysis of the cell lines for the presence of tat protein by transfecting HIV-long terminal repeat (LTR) linked to chloramphenicol acetyl transferase (CAT) revealed low, moderate, and high tat producer cell lines. RD-tat cell lines also showed enhanced virus production upon transfection of HIV-1 proviral DNA. Further, tat producer cell lines showed a high amount of hybrid virus in comparison to the control RD cells upon transfection of truncated viral DNAs. Thus, RD-tat cell lines would be valuable target cells for generating homogeneous viruses upon transfection of viral DNA.
Velpandi
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Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104.
Monken
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Srinivasan
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D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004279:DNA, Viral / Q000032:analysis; D015683:Gene Products, gag / Q000235:genetics; D015756:Genes, tat; D016655:HIV Core Protein p24; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D011533:Proviruses / Q000235:genetics; D011995:Recombination, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D012208:Rhabdomyosarcoma / Q000235:genetics*; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000737:chemistry*; D014758:Viral Core Proteins / Q000235:genetics
D004279:DNA, Viral; D015683:Gene Products, gag; D016655:HIV Core Protein p24; D014758:Viral Core Proteins; D015500:Chloramphenicol O-Acetyltransferase