Phyllosphere QS receptors C14:1 and C14:2 labeling 1
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ABSTRACT: Extracted cultures of phyllosphere bacteria's QS receptors (heterologously expressed in E. Coli) the have been incubated with an a,b-unsaturated QS signal
Project description:Biofilm formation by Escherichia coli was significantly inhibited when co-cultured with Stenotrophomonas maltophilia in static systems. Genes of E. coli involved in species interactions with S. maltophilia were identified in order to allow the study of the mechanisms of inhibited E. coli biofilm formation in co-culture. A total of 89 and 108 genes were identified as differentially expressed in mixed species cultures when growing as biofilm and as planktonic cultures, respectively, compared to the counterpart of pure cultured E. coli. Differential expression of certain identified genes was confirmed using E. coli reporter strains combined with single-cell based flow cytometry analysis. Co-culture with S. maltophilia affected genes involved in metabolism, signal transduction, cell wall composition, and biofilm formation of E. coli. Several selected genes were further confirmed as affecting E. coli biofilm formation in mixed species cultures with S. maltophilia. The data suggest that these genes were involved in species interactions between E. coli and S. maltophilia. This SuperSeries is composed of the SubSeries listed below.
Project description:We synthesize a synthetic representation of the human serine phosphoproteome, expressed heterologously as >=31 amino acid phosphopeptides in an engineered strain of E. coli. We confirm that tens of thousands of these phosphopeptides can be successfully expressed and retain important functional characteristics of serine phosphorylation as it occurs in native eukaryotic systems.
Project description:This dataset includes samples of the Maculalctone biosynthetic gene cluster (mac BGC) captured in Nodularia sp. NIES-3585 and heterologously expressed in E. coli.
Total = complete BGC
CDEF = minimal BGC
Empty vector = control
Project description:Native top-down analysis via ETnoD and HCD on the Orbitrap Eclipse of SARS-CoV-2 nucleocapsid (N) protein heterologously expressed in E. coli
Project description:KrÜppel-like factor 10 (KLF10), deficient in two thirds of 105 resected pancreatic adenocarcinoma (PDAC) patients, was associated with rapid loco-regional recurrence and large tumor size. Additional KLF10 depletion in KC (LSL: KrasG12D; Pdx1-CRE) mice accelerated progression from pancreatic intraepithelial neoplasia to PDAC. In Panc-1 cells with KLF10 deficiency (Panc-1-pLKO-shKLF10), increased sphere formation, stem cell markers expression, and tumor growth were noted compared with those of control. Over-expressing KLF10 genetically or pharmacologically using metformin, reversed the stem cell phenotypes induced by KLF10 depletion. Ingenuity pathway analysis and gene set enrichment analysis showed Notch signal molecules including Notch receptors 3 and 4 were over-expressed in Panc-1-pLKO-shKLF10. KLF10 transcriptionally suppressed Notch 3 and 4 receptors by competing with ELF3, a positive regulator, for promoter binding. Downregulating Notch signal genetically or pharmacologically ameliorated stem cell phenotypes of Panc-1-pLKO-shKLF10. Elevating KLF10 expression by metformin with concomitant evodiamine, a non-toxic Notch 3 methylation stimulator, delayed murine tumor growth of PDAC with KLF10 deficiency without prominent toxicity. These results demonstrated a novel signal pathway of KLF10 in modulating stem cell phenotypes of PDAC via transcriptional regulating Notch signal pathway. Elevating KLF10 and suppressing Notch signal may cooperatively reduce PDAC tumorigenesis and malignant progression.
Project description:Analysis of Archangium gephyra and 2 heterologous strains of E. coli expressing AHLs synthases AgpI and VitI. Control samples include E. coli strain and E. coli strain + empty expression plasmid pET28b. We determine myxobacterial AHL synthases produce acylhomoserine lactones C8-AHL and C9-AHL when heterologously expressed. No AHL metabolites observed in A. gephyra extracts.
Project description:We perform differential RNA-seq comparing a treatment with 1.4% saccharin to a mock treatment in E. coli K12 in order to describe the alterations in global transcription produced by this artificial sweetener. Three biological replicates of saturated E. coli K12 cultures were diluted 1/100 in 25 mL LB medium (in duplicate, to obtain 3 treated cultures and 3 mock controls). The cultures were grown (37 ºC, 180 rpm) to approximately OD600 0.3 and they were treated with a final concentration of 1.4% saccharin or a water control. The cultures were further incubated until reacing OD600 0.8. At this point, cells were harvested, treated with RNAlate for preservation of total RNA and stored at -80 C. After that, total RNA was extracted from each sample. As a result, 305 genes appeared upregulated by the saccharin treatment, whereas 419 were downregulated.
Project description:Arthrobacter chlorophenolicus A6 is a 4-chlorophenol degrading soil bacterium with high phyllosphere colonization capacity. Till now the genetic basis for the phyllosphere competency of Arthrobacter or other pollutant-degrading bacteria is uncertain. We investigated global gene expression profile of A. chlorophenolicus grown in the phyllosphere of common bean (Phaseolus vulgaris) compared to growth on agar surfaces.
Project description:Highly polymorphic allorecognition systems have been characterized in numerous invertebrate species, and exhibit discriminatory capabilities reminiscent of vertebrate adaptive immunity. As these systems utilize germline encoded receptors, the mechanisms underlying allelic discrimination are unknown. The invertebrate chordate, Botryllus schlosseri, undergoes a natural transplantation reaction controlled by a highly polymorphic, polygenic locus (called the fuhc) with over 1,000 allelic haplotypes found worldwide. Two individuals are compatible if they share one or both fuhc alleles, and we had found that polymorphic discrimination is due to the integration of signals from two allorecognition receptors encoded within the fuhc locus, called fester and uncle fester. Here we show that these two receptors are members of an extended family consisting of >35 genes, now called the Fester family (FF), and co-expressed with members of another diverse gene family, the fester co-receptors (FcoR). Both FF and FcoR are Immunoglobulin superfamily members and each FcoR encodes conserved tyrosine signal transduction motifs, including ITIMs or hemITAMs. FF and FcoR are expressed and encoded as cognate pairs in two polymorphic haplotypes: one within the fuhc locus, and another on a separate chromosome, and remarkably, copy number variation between haplotypes is of gene pairs. Furthermore, two FcoR genes can swap ITIMs and hemITAMs by alternative splicing, suggesting that dynamic tuning of activating and inhibitory signaling is required for allelic discrimination. These results indicate that conserved signal processing mechanisms are the foundation of both allelic discrimination in Botryllus, and recurring convergent evolution of allorecognition receptors observed from invertebrates to mammals.