Shotgun Proteomic Analysis of a Shotgun Proteomic Analysis of a Model 7-Member Human Gut Microbiota created in Gnotobiotic Mice

摘要

Preliminary data shows great promise for this technique, with excellent technical reproducibility for all 7 microbial strains and proteins identified We have applied shotgun proteomics to 3 mouse cecal samples and the mass spectrometric results match the inoculated microbial species distributions as expected Most abundant identified proteins from the host and microbes included common metabolic proteins, ribosomal proteins, and proteins involved in energy metabolism and digestion Improvements are necessary to generate reproducible and efficient lyses and protein extraction between samples, as well as increase the depth of proteome measurements. Future work will be directed at optimizing the lysis method for cecal material. Lysis methods such as bead beating, sonication, and repetitive freeze/thaw cycling will be tested. These results emphasize the value of integrating gnotobiotics with high resolution proteomics as a strategy for developing the experimental and computational pipeline needed to characterize gene expression in more complex, human body habitatassociated microbial communities.