Studies to Identify Differences between Brucella abortus Vaccine and Virulent Strains
Abstract
This project consisted of three phases, each of which was designed to provide insight into the genetic basis for the difference in virulence between strains of Brucella abortus. In the first phase plasmids were constructed which contain portions of the genes encoding the porin proteins of E. coli (OMP F and OMP C). The second phase involved an attempt to isolate the plasmid M13. This would be used as a cloning vector to subclone B. abortus fragments in order that they may be sequenced by the Exonuclease III method. Finally, the third phase was an antibiotic resistance study. This allowed the determination of the differences in resistance to various antibiotics between several strains of B. abortus.
In the design of plasmids containing the OMP C and OMP F gene fragments, the fragments were subcloned into pBR322. These were then transfected into HB101 and amplified. The plasmids were then isolated and used in nick translation for probing B. abortus DNA.
The next phase involved the isolation of the plasmid M13. Following the infection of JM101 host cells with M13 phage, the cells were allowed to amplify the M13 plasmid. The plasmid was then recovered by the SDS method and purified over cesium chloride. It was then stored for use for sequencing by Exonuclease III.
Finally, the antibiotic resistance study involved streaking the B. abortus strains 19, 2308, and RE51 on increasing antibiotic concentrations. It was found that there was a significant difference in the resistance to chloramphenicol, tetracycline, and spectinomycin.
Description
Program year: 1986-1987Digitized from print original stored in HDR
Citation
Lancaster, Brent (1987). Studies to Identify Differences between Brucella abortus Vaccine and Virulent Strains. University Undergraduate Fellows. Available electronically from https : / /hdl .handle .net /1969 .1 /CAPSTONE -LancasterB _1987.