A REPORTER SYSTEM USING A FLOW CYTOMETER TO DETECT PROMOTER ENHANCER ACTIVITY IN LYMPHOID-CELL LINES/

We have devised an experimental system using a flow cytometer to exami ne the promoter/enhancer activity of DNA fragments in human lymphoid c ell lines. Murine CD8 alpha gene cDNA used as a reporter gene was inse rted in the reporter constructs under the control of various promoter/ enhancers. Furthermore, the Epstein-Barr virus (EBV) OriP, which suppo rts a high transient expression, was also included in the reporter con structs. Cell lines expressing EBV nuclear antigen-1 (EBNA-1) were tra nsfected with the reporter constructs by electroporation. The expressi on of the reporter gene was measured by a flow cytometric analysis. Th is experimental system is quite simple and may be especially useful fo r the analysis of transcriptional elements functioning in lymphocytes.