Cytochemical analysis of human T cell leukaemia virus I LTR-regulated beta-galactosidase gene expression using a novel integrated cell system.
To develop a reporter system to study the response of an integrated retroviral LTR and cellular and viral events which influence transcription, the 5' LTR of HTLV-1 was coupled to the Escherichia coli beta-galactosidase gene (lacZ). This construct was assembled within a vector containing the neomycin resistance gene controlled by the SV40 promoter, and introduced into HeLa cells. Expression from the LTR in one clone was upregulated by positive regulators of HTLV-1 expression, including 12-O-tetradecanoylphorbol-13-acetate (TPA) and the HTLV-1 transregulatory protein (tax), as has been previously reported using transient transfection assays. This method proved to be a rapid and reproducible assay for the measurement of integrated viral LTR activation in a single cell system.
Copeland
K F
KF
Department of Chronic and Infectious Diseases. TNO-MBL, Rijswijk, The Netherlands.
Haaksma
A G
AG
Derse
D
D
Goudsmit
J
J
Heeney
J L
JL
D004926:Escherichia coli / Q000235:genetics; D005069:Evaluation Studies as Topic; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D016356:Gene Products, tax / Q000494:pharmacology; D017930:Genes, Reporter; D005814:Genes, Viral; D005822:Genetic Vectors; D006367:HeLa Cells; D006651:Histocytochemistry; D015368:Human T-lymphotropic virus 1 / Q000201:enzymology* / Q000235:genetics*; D006801:Humans; D007763:Lac Operon; D012091:Repetitive Sequences, Nucleic Acid; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014773:Virology / Q000379:methods; D001616:beta-Galactosidase / Q000235:genetics*
D016356:Gene Products, tax; D001616:beta-Galactosidase; D013755:Tetradecanoylphorbol Acetate