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Next Generation Sequencing of bacterial surface-displayed ENAH EVH1 ligand peptides after a FACS titration sort (MassTitr)

ABSTRACT: Next Generation Sequencing of bacterial surface-displayed ENAH EVH1 ligand peptides after a FACS titration sort (MassTitr)

PROVIDER: PRJNA702444 | ENA |

REPOSITORIES: ENA

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Next Generation Sequencing of bacterial surface-displayed ENAH EVH1 ligand peptides after a FACS titration sort (MassTitr)

Project description:We used bacterial surface display and FACS to sort peptide binders to a tetramerized ENAH EVH1 domain at multiple concentrations of ENAH into 4 gates. This experiment was repeated three times. By calculating the distribution of each clone across gates at multiple concentrations, we are able to assess whether a clone exhibited concentration-dependent binding to ENAH EVH1 domain, allowing us to distinguish binders from nonbinders.

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Project description:The metabolic status of individual cells in microbial cultures can differ being relevant for biotechnology, environmental and medical microbiology. However, it is hardly understood in molecular detail due to limitations of current analytical tools. Here, we demonstrate that FACS in combination with proteomics can be used to sort and analyze cell populations based on their metabolic state. A previously established GFP reporter system was used to detect and sort single Corynebacterium glutamicum cells based on the concentration of branched chain amino acids (BCAA) using FACS. A proteomics workflow optimized for small cell numbers was used to quantitatively compare proteomes of a ?aceE mutant, lacking functional pyruvate dehydrogenase (PD), and the wild type. About 800 proteins could be quantified from 1,000,000 cells. In the ?aceE mutant BCAA production was coordinated with upregulation of the glyoxylate cycle and TCA cycle to counter the lack of acetyl CoA resulting from the deletion of aceE.

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In-culture titration experiment for sRSE

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In-culture titration experiment for sRSE

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DNA methylation titration data from two samples with known methylation differences

Project description:Two samples with known methylation differences. Sample A is NA 10923 from Coriell Institute for Medical Research. It is a B-Lymphocyte sample from a male donor. Sample B is HTB-38 cell line from ATCC (http://www.atcc.org). It is a colon cancer sample from a female donor. Sample A and B were normalized into the same concentration, and then mixed in five different titration ratios: 100:0, 90:10, 75:25, 50:50 and 0:100 Sample A and Sample B were mixed in five different titration ratios: 100:0, 90:10, 75:25, 50:50 and 0:100

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FACS sorted podocytes, mesangial cells

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The IDA-LIKE peptides IDL6 and IDL7 are negative modulators of stress responses in Arabidopsis thaliana

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