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 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.
Mazda
O
O
Department of Immunology, Kyoto University, Japan.
Teshigawara
K
K
Fujimoto
S
S
Hattori
N
N
Dou
Y M
YM
Katsura
Y
Y
D001402:B-Lymphocytes / Q000276:immunology*; D001483:Base Sequence; D015139:Blotting, Southern; D016827:CD8 Antigens / Q000235:genetics*; D002460:Cell Line; D002467:Cell Nucleus / Q000276:immunology; D004247:DNA / Q000235:genetics; D004742:Enhancer Elements, Genetic; D005434:Flow Cytometry / Q000379:methods*; D015870:Gene Expression; D004854:Herpesvirus 4, Human / Q000276:immunology; D006801:Humans; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D015375:Receptors, Interleukin-2 / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology*; D014158:Transcription, Genetic; D014162:Transfection
D016827:CD8 Antigens; D015375:Receptors, Interleukin-2; D004247:DNA