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Rapid Synthesis and Screening of Chemically Activated Transcription Factors with GFP-based Reporters
Published on: November 26, 2013
An improved growth hormone reporter gene shows a different pattern of transactivation by promoter fragments
Pierre Schembri-Wismayer1, George D Birnie
1Beatson Institute for Cancer Research, Scotland, Glasgow, UK. pierre.shembri-wismayer@um.edu.mt
Abstract:
Granulocytic differentiation of myeloid NB4 cells was noted to transactivate the genomic human growth hormone (hGH) reporter construct in the absence of a driving test promoter. Such spurious hGH transactivation also occurred upon co-transfection of plasmids expressing the transactivators c-Myb and C/EBPalpha into undifferentiated NB4 cells. Modifications to the reporter gene and the vector resulted in a reporter construct unaffected by such stimuli. When human neutrophil peptide 1 (hnp-1) gene promoter fragments were tested using both the original and modified reporter constructs, a clear difference in the pattern of transactivation was noted. This suggests that promoter and enhancer elements in the reporter construct plasmid DNA can effect the analysis of the test promoter.
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