A reporter system using a flow cytometer to detect promoter/enhancer activity in lymphoid cell lines

J Immunol Methods. 1994 Feb 28;169(1):53-61. doi: 10.1016/0022-1759(94)90124-4.

Abstract

We have devised an experimental system using a flow cytometer to examine the promoter/enhancer activity of DNA fragments in human lymphoid cell lines. Murine CD8 alpha gene cDNA used as a reporter gene was inserted in the reporter constructs under the control of various promoter/enhancers. Furthermore, the Epstein-Barr virus (EBV) OriP, which supports a high transient expression, was also included in the reporter constructs. Cell lines expressing EBV nuclear antigen-1 (EBNA-1) were transfected with the reporter constructs by electroporation. The expression of the reporter gene was measured by a flow cytometric analysis. This experimental system is quite simple and may be especially useful for the analysis of transcriptional elements functioning in lymphocytes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • B-Lymphocytes / immunology*
  • Base Sequence
  • Blotting, Southern
  • CD8 Antigens / genetics*
  • Cell Line
  • Cell Nucleus / immunology
  • DNA / genetics
  • Enhancer Elements, Genetic*
  • Flow Cytometry / methods*
  • Gene Expression
  • Herpesvirus 4, Human / immunology
  • Humans
  • Molecular Sequence Data
  • Promoter Regions, Genetic*
  • Receptors, Interleukin-2 / genetics
  • T-Lymphocytes / immunology*
  • Transcription, Genetic
  • Transfection

Substances

  • CD8 Antigens
  • Receptors, Interleukin-2
  • DNA