Fig. 6: Using RT-IVT assays to detect the DNA-binding coefficient of transcriptional inhibitor.
From: RT-IVT method allows multiplex real-time quantification of in vitro transcriptional mRNA production
a Principles for DNA template design. Figure was created with Biorender (www.bioender.com). b The relative quantitative results (signal corrected) for the DNA templateFur after Zn1Mn1Fur was added. The reaction configuration and detection conditions are identical to those shown in Fig. 3a, except 30 nM DNA templateFur replaced DNA template 1, and 0‒2 µM Zn1Mn1Fur was added. c The KD value for Fur binding to the Fur box DNA. The initial transcription velocity for the 0‒2 µM Zn1Mn1Fur added groups were measured by fitting the slope of the 1‒15 min reaction region of Fig. 6b. The transcriptional repression rates were calculated using the initial transcription velocity of 0 µM Zn1Mn1Fur group subtracted by the corresponding groups. The Zn1Mn1Fur concentration corresponds to half of the maximum transcriptional inhibition rates and represents the KD value of Zn1Mn1Fur with the Fur box DNA. The values represent the mean ± standard deviation of n = 2 replicates.
