Project Number: 3625-32000-061-03
Project Type: Specific C/A

Start Date: Sep 21, 2001
End Date: Aug 28, 2004

Objective:
To construct libraries of DNA from B. abortus in vectors. To analyze the libraries to show that they are random and that they have a high percentage of clones with inserts for use in a genomic sequencing project. To prepare plasmid DNA from the libraries that can be used as templates for DNA sequence. To determine by use of automated DNA sequencing the sequence of B. abortus. To align, annotate, and analyze the sequence for genes in order to understand the metabolic capabilities and identify virulence factors.

Approach:
The DNA will be isolated from bovine isolate. The DNA will be sheared and electorphoresed to obtain fragments of 2 to 2.5 kb. After the ends of the sheared isolate fragments are repaired, the fragments will be ligated into the linearized phosphatased vector pUC 18. The vector library will be used to transform E. coli and 24,000 plasmid templates will be isolated and sequenced at the University of Minnesota. A portion of the plasmid templates isolated at the University of Minnesota will be sent to NADC for sequence analyses. The sequence files will be aligned by use of software to assemble the genomic sequences. The sequence will be analyzed by use of public domain computer software to determine open reading frames, the homology of the open reading frames to bacterial databases. The linkage of the open reading frames and the probable function of the open reading frames will be noted.

Project Team

Halling, Shirley Vivek Kapur - Associate Professor (515)663-7370

Project Annual Reports

FY 2003   FY 2002

Related National Programs

Animal Health (103)