ABSTRACT: Mass screening of crude extracts prepared from Actinokineospora auranticolor fermentation and LC-MSMS data acquisition for structure analysis.
Project description:Mass screening of crude extracts prepared from Actinokineospora auranticolor fermentation and LC-MSMS data acquisition for structure analysis.
Project description:This project aims to analyse the proteins of outer membrane vesicles (OMVs) released by a Serratia bacteria strain Su_YN1, which is isolated from field-caught mosquito gut. Su_YN1 produces massive OMVs under host serum induction. The data contain LC-MSMS results of the OMVs released without serum induction (WT-), and serum induced OMVs of WT strain (WT+) and several gene disruption strains (disruption of the ABC transporter genes ABC1 and ABC2). This project was conducted by CEMPS lab and Shanghai hoogen biotech co.ltd. The CEMPS lab prepared the purified OMV samples and Shanghai hoogen biotech co.ltd conducted the LC-MSMS analysis.
Project description:We constructed genetically modified Eimeria acervulina that expressed VP2 protein, a protective antigen from infectious bursal disease virus (IBDV), on the surface or in the microneme of sporozoites. we identified the transgenic E. acervulina expressing VP2 by RT-qPCR,LC-MSMS and Werstern Blot.
Project description:With the optional setting of multiple stepped collisional energies (NCEs), higher-energy collisional dissociation (HCD) as available on Orbitrap instruments is a widely adopted dissociation method for intact N-glycopeptide characterization, where peptide backbones and N-glycan moieties are selectively fragmented at high and low NCEs, respectively. Initially, a dependent setting of a central value plus minus a variation is available to the users to set up NCEs, and the combination of 30±10% to give the energies 20%/30%/40% has been mostly adopted in the literature. With the recent availability of independent NCE setup, we found that the combination of 20%/30%/30% is better than 20%/30%/40%; in the analysis of complex intact N-glycopeptides enriched from gastric cancer tissues, total IDs with spectrum-level FDR≤1%, site-specific IDs with site-determining fragment ions and structure-specific IDs with structure-diagnostic fragment ions were increased by 42% (4,767->6,746), 57% (599->942), and 97% (1771->3495), respectively. This finding will benefit all the coming N-glycoproteomics studies using HCD as the dissociation method.
Project description:Enzymatic Methyl-seq (EM-seq) is an enzyme-based method giving us reliable methylome data from small amounts of purified DNA. However, it is unclear whether EM-seq library can be constructed from crude cell lysate containing genomic DNA. We evaluated the quality of EM-seq libraries directly prepared from crude cell lysate.