炭疽感染的蛋白质组学课件.pptVIP

Criteria Target protein is present during infection. Target protein’s mRNA is also expressed. Target protein has close homology to known essential factors, or virulence factors, from other organisms. Target protein maintains critical activities, regions or structures that are non-mammalian. B. anthracis morphologies Bacillus Life Cycle B. anthracis genome Chromosome 5.23 MB 5508 genes pXO1 toxin plasmid 182 KB 217 genes pXO2 capsule plasmid 95 KB 113 genes B. anthracis genome—new possibilities DNA microarray construction for RNA expression studies Mass Spectrometry-based shotgun proteomics Sporulation in B. anthracis Cluster analysis of growth phase-regulated B. anthracis genes Growth phase dependent expression occurs in five waves B. anthracis genome—new possibilities DNA microarray construction for RNA expression studies Mass Spectrometry-based shotgun proteomics Expression of the B. anthracis spore proteome Expression of the B. anthracis spore proteome B. anthracis Germinant Sensors Tri-cistronic operons Expressed late during sporulation Disruption leads to a germinant specific defect in germination Chromosomal: gerH gerS gerL gerK gerY* gerA* pXO1: gerX B. anthracis Germinant Sensors Bacillus Life Cycle Target Validation #1 Determine the contributions made by each putative germinant sensor to the germination profile of B. anthracis endospores. Disruption of Germinant Sensors Disruption of Germinant Sensors Germination Analysis Germination Assays D-gerL D-gerS D-gerH Germinant Recognition Macrophage Associated Germination Target Validation #2 dltA, is the first of four genes in the dlt operon, which controls the incorporation of D-alanine into lipoteichoic acids on the cell surface. Essential for: maintenance of normal cell shape, control of autolytic enzymes, and resistance to cationic anti-microbial peptides in Gram positive organisms. Loss of dlt operon has been shown to decrease the virulence of S. aureus and L. monocyto