Mazda O, Teshigawara K, Fujimoto S, Hattori N, Dou Y M, Katsura Y
Department of Immunology, Kyoto University, Japan.
J Immunol Methods. 1994 Feb 28;169(1):53-61. doi: 10.1016/0022-1759(94)90124-4.
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.
