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For the BEAROFF device, the C. jejuni repressor CmeR was fused to the VP16 transactivation domain to create a synthetic transactivator that activates minimal promoters containing tandem operator modules (Ocme) in a bile acid-repressible manner.
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Since the parameters that could be varied are the levels of Lac I and the number of tandem operators, we plotted f (E) as a function of these two variables using equations 3 and 5.
For promoters harboring one, two, and eight tandem operators, corresponding increases in maximal expression were observed (indicating a possible increase in vertical extension, although full response curves were not presented); interestingly, a less-obvious decrease in leakage was also seen.
Binding of TetR-GFP to tandem TetO operator sites in the tag creates a bright fluorescent dot that can be tracked in 3D using machine vision software [31], [32].
To investigate the effect on sister chromatid cohesion, we used a strain in which the URA3 locus is marked by the binding of Tet repressor-GFP to multiple tandem Tet operators (Michaelis et al, 1997).
An EBV-derived plasmid (pLON-33K) containing oriP, 264 copies in tandem of Lac operator (LacO), and a neomycin-resistance gene was introduced into and established in clones of HeLa cells stably expressing EBNA1 (HeLa-EBNA1).
Try it yourself on a tandem flight with local operator Millau Evasion.
The IRF-1 gene was cloned into the expression vector pcDNA™4/TO/myc-His under strong CMV promoter with two copies of the Tet operator in tandem (TetO2) in the promoter.
To investigate this cohesion defect, we monitored splitting of URA3 sequences, which were visualized by the binding of Tet repressor-GFP to a tandem array of Tet operators (Michaelis et al, 1997).
pFX92 with two cassettes of 120 tandem copies of the tetO operator separated by a kan gene.
This allele has a GmR cassette between two arrays of 120 tandem repeats of the tetA operator site.
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