Accession Number:

ADA625389

Title:

Rapid Identification of Bacterial Virulence Factors

Descriptive Note:

Final status rept. 1 Feb 2012-15 Apr 2014

Corporate Author:

VITAL PROBES MAYFIELD PA

Personal Author(s):

Report Date:

2014-04-15

Pagination or Media Count:

59.0

Abstract:

The main objective of this work is to demonstrate that Rapid Virulence Factor Discovery System RVFDS can be used to identify virulence factors of MRSA and Brucella abortus. This will be accomplished by constructing an MRSA and B. abortus random expression library REL in E. coll. The MRSA REL contains hundreds of potential virulence factors. Putative virulence factors that interacted with the ECM components fibronectin, laminin, and plasminogen, collagen, and anti-MRSA polyclonal antibodies will be eluted from pull-down columns that contain these bait proteins. Putative MRSA virulence factors will be identified by LC-MSMS. Additional virulence factors will be identified using a wider range of ECM components as bait proteins. In the Phase I investigation the average size of the REL insert was in the 1.5 to 2.0kb range. In Phase II the average size of the insert will be increased to 3.0kb in order to decrease redundancy. Larger inserts and REL protein sizes will also facilitate LC-MSMS identification and shorten the time needed for data analysis. Potential proteins that are discovered in Phase II work may serve as targets for future vaccines, antimicrobials, and therapeutic measures.

Subject Categories:

  • Microbiology

Distribution Statement:

APPROVED FOR PUBLIC RELEASE