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Constitutive expression of heterologous genes is a key step for metabolic engineering of heterogeneous microbial cells. Screening of suitable promoters is often laborious and time-consuming. Here we describe a high-throughput strategy based on the analysis of the transcriptional response of a broad panel of promoters to the addition of the positively charged agent cetyltrimethylammonium bromide (CTAB) to culture medium. The CTAB-evoked responses of the promoters in Pseudomonas putida KT2440 were quantified through an alkaline phosphatase (phoA) assay and the promoter activities were scored based on the concentration of the CTAB required to induce detectable levels of alkaline phosphatase. The frequency distribution of the activity scores enabled us to estimate the mean promoter activity and the mean promoter activity of the strongest-expressed promoters. Furthermore, a cross-referencing with a database of known promoters provided a measure of the promoter strength. Using this strategy, we screened 51 promoters from different origins to select those with the highest mean promoter activity and the highest mean promoter activity of the strongest-expressed promoters. A summary of the promoter activity analysis and our selection of promoters for constitutive expression in P. putida KT2440 is provided. This strategy could be applied to other bacteria for the constitutive expression of heterologous proteins.This is a pretty big change and is a change for the better. Even though the ability to use way more characters than ever was troubling to some, it is an overall improvement. The main problem is not having more characters available in tournament play (as the guild is not very large, we can’t play that many), but having more than one character in the friend list at a time.

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Víctor M. Cruz (born February 8, 1972) is a Cuban politician and diplomat, and the current Permanent Representative of Cuba to the United Nations since 2019.

Early life
Víctor M. Cruz was born on February 8, 1972 in Havana, Cuba. He graduated from the University of Havana as an engineer, and obtained a degree in international relations from the Central University of Venezuela.

Diplomatic career

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