Document 0831
 DOCN  M9590831
 TI    Real-time analysis of the transcriptional regulation of HIV and hCMV
       promoters in single mammalian cells.
 DT    9509
 AU    White MR; Masuko M; Amet L; Elliott G; Braddock M; Kingsman AJ; Kingsman
       SM; Amersham International, Bucks, UK.
 SO    J Cell Sci. 1995 Feb;108 ( Pt 2):441-55. Unique Identifier : AIDSLINE
       MED/95286692
 AB    The regulation of human cytomegalovirus (hCMV) and human
       immunodeficiency virus (HIV) gene expression has been studied in single
       intact mammalian cells. Viral promoters were placed upstream of the
       firefly luciferase reporter gene and the resulting hybrid reporter
       constructs were stably integrated into the HeLa cell genome. A highly
       sensitive photon-counting camera system was used to study the level of
       gene expression in single intact cells. Luciferase expression was
       studied in the absence of activators of viral gene expression, in the
       presence of the HIV-1 TAT transactivator protein, or in the presence of
       sodium butyrate, a non-viral activator of gene expression. In the
       absence of any activator of gene expression, while expression was
       undetectable in most cells, significant levels of basal luciferase
       activity were observed in a few cells, indicating heterogeneity in gene
       expression in the cell population. In the presence of the general
       activator of viral gene expression, sodium butyrate, transcriptional
       activation from the viral promoters gave rise to significant and
       relatively homogeneous levels of luciferase expression in a majority of
       cells. The luciferase imaging technology was used for the real-time
       analysis of changes of gene expression within a single cell. This
       non-invasive reporter assay should become important for studies of the
       temporal regulation of gene expression in single cells.
 DE    Animal  Beetles/ENZYMOLOGY  Butyric Acids/PHARMACOLOGY
       Cytomegalovirus/*GENETICS  Gene Expression Regulation, Viral/DRUG
       EFFECTS  Gene Products, tat/GENETICS  Hela Cells  Human  HIV-1/*GENETICS
       HIV-2/*GENETICS  Image Processing, Computer-Assisted
       Luciferase/GENETICS  *Promoter Regions (Genetics)  Support, Non-U.S.
       Gov't  Time Factors  *Transcription, Genetic  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).