Approximately one third of the human population (~2 billion persons world-wide) harbor viable tubercle bacterium in their lungs, and almost 2 million of these die annually from tuberculosis (TB). The HIV pandemic, the rise of antibiotic-resistant strains of Mycobacterium tuberculosis (MTB) and increased global migration threaten to push these rates even higher. Innovative methods to combat this ancient pathogen are sorely needed. This proposal describes how the personnel will combine their unique expertise in modern computational, statistical and molecular techniques to build a "whole genome" view of MTB gene expression. The Primary Investigator's existing database of DNA microarray experiments and statistical methods will be used to build a transcript map for MTB, incorporating the co-expression of potential operon gene pairs. RT-PCR and primer extension will be used to refine this predicted map, which will then be published via peer review, posted on the internet and distributed to major biological databases. This project will expand our understanding of genome structure and transcription in MTB, an important step in understanding how this bacterium adapts and remains pathogenic. Undergraduate student researchers mentored by PI Rutherford and Dr. Roback will receive interdisciplinary training at the intersections of genetics, statistics and bioinformatics, and the research infrastructure at two institutions will be enhanced. Specific Aims: 1. Enhance the transcript map by incorporating comparative and pathway-based methods of operon prediction. 2. Test the map by expanding our verification of predicted transcripts in the laboratory. 3. Deliver an accurate and useable MTB transcript map to the scientific community. Given the recent upward pressures in tuberculosis (TB)'s annual death toll of 2 million people, innovative approaches to treatment are sorely needed. This proposal uses a combination of modern computational, statistical and molecular techniques to discover how the pathogenic bacterium that causes TB organizes and uses its genome to cause disease. Undergraduate students will be vital participants in this research, receiving extensive exposure to contemporary, interdisciplinary scientific research. [unreadable] [unreadable] [unreadable]