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The correct promoter insert was confirmed for the plasmids via PCR.
Ideally, the findings generated using all of these published models should be reconfirmed in systems that use the correct promoter.
ReLA is able to successfully discriminate the correct promoter region, including sites that have been experimentally proven (Blanco et al., 2006a).
In addition to not using the correct promoter to test enhancer elements, the choice of cell type could influence the results for enhancers, which function in a highly cell-type specific manner.
Notably, a recent study suggests that using the correct promoter (referred to as UPKII) to create mice expressing the Simian virus 40 (SV40) large T-antigen (which inactivates both p53 and Rb) results in the development of CIS only, with activation of pro-angiogenic factors but not progression to invasive cancer (Ayala de la Pena et al., 2011).
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In particular, rapid and accurate localization and binding of correct transcription promoter sites by the RNA polymerase (RNAP) holoenzyme may be one of the critical steps limiting the overall efficiency of gene expression, especially in bacteria, where genomic DNA is not protected by nucleosome-like structures.
However, as noted above, it was more recently shown that urothelial targeting of SV40 using the correct UPKII promoter results in CIS without progression to invasive disease (Ayala de la Pena et al., 2011).
For correct expression, promoters such as the cauliflower mosaic virus (CaMV) 35S promoter (P35S) are needed.
Furthermore, the conservation of quantitative responses among orthologous Msx1 promoters to the same input confirms our selection of the correct fugu Msx1 promoter region.
Animal studies confirm that correct regulation of promoter proximal pausing is critical for development and health in adult life.
So if the model of small family-farms is more politically correct to the promoters of fair trade, then Brazilian coffee should not be excluded from their marketing efforts.
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