Title page for ETD etd-10022003-120410

Type of Document Master's Thesis
Author Sturgill, David Matthew
Author's Email Address dsturgil@vt.edu
URN etd-10022003-120410
Title Comparative Genome Analysis of Three Brucella spp. and a Data Model for Automated Multiple Genome Comparison
Degree Master of Science
Department Biology
Advisory Committee
Advisor Name Title
Gibas, Cynthia J. Committee Chair
Boyle, Stephen M. Committee Member
Hilu, Khidir W. Committee Member
Melville, Stephen B. Committee Member
Weller, Jennifer W. Committee Member
  • Brucella
  • Comparative Genomics
  • Bioinformatics
  • Host-pathogen interaction
Date of Defense 2003-09-12
Availability unrestricted
Comparative analysis of multiple genomes presents many challenges ranging from management of information about thousands of local similarities to definition of features by combination of evidence from multiple analyses and experiments. This research represents the development stage of a database-backed pipeline for comparative analysis of multiple genomes. The genomes of three recently sequenced species of Brucella were compared and a superset of known and hypothetical coding sequences was identified to be used in design of a discriminatory genomic cDNA array for comparative functional genomics experiments. Comparisons were made of coding regions from the public, annotated sequence of B. melitensis (GenBank) to the annotated sequence of B. suis (TIGR) and to the newly-sequenced B. abortus (personal communication, S. Halling, National Animal Disease Center, USDA).

A systematic approach to analysis of multiple genome sequences is described including a data model for storage of defined features is presented along with necessary descriptive information such as input parameters and scores from the methods used to define features. A collection of adjacency relationships between features is also stored, creating a unified database that can be mined for patterns of features which repeat among or within genomes.

The biological utility of the data model was demonstrated by a detailed analysis of the multiple genome comparison used to create the sample data set. This examination of genetic differences between three Brucella species with different virulence patterns and host preferences enabled investigation of the genomic basis of virulence. In the B. suis genome, seventy-one differentiating genes were found, including a contiguous 17.6 kb region unique to the species. Although only one unique species-specific gene was identified in the B. melitensis genome and none in the B. abortus genome, seventy-nine differentiating genes were found to be present in only two of the three Brucella species. These differentiating features may be significant in explaining differences in virulence or host specificity. RT-PCR analysis was performed to determine whether these genes are transcribed in vitro. Detailed comparisons were performed on a putative B. suis pathogenicity island (PAI). An overview of these genomic differences and discussion of their significance in the context of host preference and virulence is presented.

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