Transcriptional activity or under control of six different promoters

Copyright information:Taken from "A novel magnetic resonance-based method to measure gene expression in living cells"Nucleic Acids Research 2006;34(6):e51-e51.Published online 5 Apr 2006PMCID:PMC1447650.© The Author 2006. Published by Oxford University Press. All rights reserved Gene activity was detected by northern blot and MR spectroscopy (MRS). () Schematic diagram of the reporter constructs used to express or . The sequences derived from the and promoters are shown as hatched and dotted rectangles, respectively. The regions included in each construct (#1–6) are indicated at the right. () Total RNA was extracted from cells growing logarithmically at 30°C in YPD and analyzed by northern blot using a probe for mRNA (upper panel). Transcription of was driven by the endogenous promoter (WT, lane 1) or heterologous promoters #1–6 (lanes 2–7). The strain was used as a negative control (lane 8) to show that the gene expression signals are specific to . The blot was probed for mRNA as a control for equal loading (lower panel). () Transcription of and in the indicated strains were measured by northern blot as described for panel (B). () P-MRS analysis of gene expression was performed using the same strains as in panel (B). Strain number is indicated to the left of each MRS scan. Peaks corresponding to MDP, inorganic phosphate (Pi) and inorganic polyP are marked at the top. MDP was added at a known concentration as a chemical shift standard. () P-MRS analysis of expression was performed as described in (D) using the same strains as in panel (). Strain number is indicated to the left of each MRS scan.