Project description:A whole genome PCR amplicon DNA microarray for B. mallei were fabricated as previously described [7]. Total RNA was isolated from in vitro cultures in LBG medium of B. mallei ATCC 23344, FMH, and JHU. The OD600 of the samples at harvest were all 0.55. The RNAs from FMH and JHU were labeled and hybridized to the array using the ATCC 23344 RNA as the reference using protocols as described. Flip-dye replicates were performed for all analyses. Two B. mallei ATCC 23344 samples grown to an O.D.600 of 1.0 on separate days and total RNA was extracted. These RNA samples were hybridized against each other as a control for the JHU vs. ATCC 23344 hybridization and the FMH vs. ATCC 23344 hybridization.
Project description:Burkholderia mallei and Burkholderia pseudomallei are both potential biological threats agents. Melioidosis caused by B. pseudomallei is endemic in Southeast Asia and Northern Australia, while glanders caused by B. mallei infections are rare. Here we studied the proteomes of different B. mallei and B. pseudomallei isolates to determine species specific characteristics. Analyzing the expressed proteomes of B. mallei and B. pseudomallei revealed differences between B. mallei and B. pseudomallei but also between isolates from the same species. Expression of multiple virulence factors and proteins of several PKS/NRPS clusters was demonstrated. Proteome analysis can be used not only to identify bacteria but also to characterize the expression of important factors that putatively contribute to pathogenesis of B. mallei and B. pseudomallei.
