An improved growth hormone reporter gene shows a different pattern of transactivation by promoter fragments.
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.
Schembri-Wismayer
Pierre
P
Beatson Institute for Cancer Research, Scotland, Glasgow, UK. pierre.shembri-wismayer@um.edu.mt
Birnie
George D
GD
D045744:Cell Line, Tumor; D015870:Gene Expression; D017930:Genes, Reporter / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D018922:HL-60 Cells; D019382:Human Growth Hormone / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D009154:Mutation; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D015533:Transcriptional Activation / Q000235:genetics*; D014162:Transfection / Q000379:methods; D015854:Up-Regulation
D011993:Recombinant Fusion Proteins; D019382:Human Growth Hormone