Project description:A popular strategy for enhancing the antibacterial properties of probiotic bacteria is to retrofit them with the ability to overproduce heterologous bacteriocins. This is often achieved from strong, non-native promoters. How the dysregulated overproduction of heterologous bacteriocins affects the fitness and antibacterial efficacy of the retrofitted probiotic bacteria is often overlooked. We conferred the prototypical probiotic Escherichia coli strain Nissle (EcN) the ability to produce different amounts of the bacteriocin microcin C (McC). Expression of the bacteriocin synthesis genes was driven from the native promoter (Pmcc-WT), or from promoters manipulated to be stronger (Pmcc-High) and weaker (Pmcc-Low) than the WT, in a plasmid-based system. Pmcc-Low and Pmcc-High retained their native regulation. A strain harbouring a non-functional promoter (Pmcc-Mut) produces no McC and was used as a control. Each strain was grown to early stationary phase, when production of McC starts, in Luria-Bertani broth at 37 degrees. The RNA was isolated and the effects of different levels of production of McC on the transcriptome of EcN was examined by RNA-Seq.
Project description:Purpose: The goal of this study are to reveal the internal mechanism of Bacillus pumilus G5 and silicon increased Glycyrrhiza uralensis Fisch. seedlings drought-tolerance by RNA-Seq. Methods: mRNA profiles of Glycyrrhiza uralensis Fisch. Seedling in five treatment: control treatment, drought stress treatment, drought stress with G5 treatment, drought stress with Si treatment and drought stress with G5 combined Si treatment. Results: The full-length transcriptome sequencing of 15 samples was completed, and the clean data of each sample was 6.28GB. All the consistent transcript sequences were aligned to the reference genome by minimap2 software and then de-redundant analysis was performed. Finally, 37267 genes were obtained. A total of 6934 DEGs were identified in four comparisons (D vs CK, DB vs D, DSi vs D, and DBSi vs D), among which are 967, 1559, 1278 and 3130 DEGs in four comparisons, respectively. Conclusions: Our study help to better understand the underlying molecular mechanisms of Bacillus pumilus G5 and silicon improve the drought-tolerance of G. uralensis.
Project description:Primary objectives: The primary objective is to investigate circulating tumor DNA (ctDNA) via deep sequencing for mutation detection and by whole genome sequencing for copy number analyses before start (baseline) with regorafenib and at defined time points during administration of regorafenib for treatment efficacy in colorectal cancer patients in terms of overall survival (OS).
Primary endpoints: circulating tumor DNA (ctDNA) via deep sequencing for mutation detection and by whole genome sequencing for copy number analyses before start (baseline) with regorafenib and at defined time points during administration of regorafenib for treatment efficacy in colorectal cancer patients in terms of overall survival (OS).