Global Proteomics Profiling of LPS Stimulated IMM cells
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ABSTRACT: Immortalized mouse macrophages (IMMs) were stimulated with 100 ng/ml lipopolysaccharide (LPS) for 0, 30, and 60 min. The samples were analyzed using LC-MS for global proteomics.
Project description:Immortalized mouse macrophages (IMMs) were stimulated with lipopolysaccharide (LPS). RNA and RNA associated proteins were UV crosslinked, and sites of RNA-association were identified using LC-MS proteomics.
Project description:To determine whether CRISPR interference can be used to recapitulate a glycosylation null mutant strain in Burkholderia cenocepacia via data independent acquisition mass spectrometry using a low level of dCas9 induction. A guideless plasmid was containing in the wildtype and pglL mutant strains, and the strains harbouring the dCas9 insertion containing a pglL-targeting plasmid Bc-pglL-C, non-target, and the guideless plasmid.
Project description:To determine the effect of CRISPR interference-mediated silencing of glycosylation in Burkholderia thailandensis harbouring a pglL-targeted guide, and a nontarget guide using Data Independent Acquisition
Project description:To determine the effect of CRISPR interference-mediated silencing of glycosylation in Burkholderia multivorans harbouring a pglL-targeted guide, and a nontarget guide using Data Independent Acquisition
Project description:To determine the affect of CRISPR interference-mediated silencing of glycosylation in Burkholderia diffusa harbouring a pglL (WL85_RS02125)-targeted guide, and a nontarget guide using Data Independent Acquisition
Project description:To determine whether CRISPR interference can be used to silence the expression of pglL in Burkholderia diffusa and therefore modulate glycosylation
Project description:To interrogate the glycoproteome following knockdown of pglL and subsequent knockdown of glycosylation in Burkholderia cenocepacia through Zwitterionic Hydrophilic Interaction Liquid Chromatography (ZIC-HILIC) glycopeptide enrichment with 0.1% rhamnose for dCas9 induction– loading control pre-enrichment. Using strains harbouring the pglL-targeting guide C (Bc-pglL-C) and guideless plasmids.