Title page for ETD etd-05202009-125845


Type of Document Master's Thesis
Author Jain, Neeta
Author's Email Address njain80@vt.edu
URN etd-05202009-125845
Title Role of entF Gene in Iron Acquisition by Brucella abortus 2308
Degree Master of Science
Department Biomedical and Veterinary Sciences
Advisory Committee
Advisor Name Title
Sriranganathan, Nammalwar Committee Chair
Boyle, Stephen M. Committee Member
Jortner, Bernard S. Committee Member
Riffle, Judy S. Committee Member
Keywords
  • colony forming units (CFU)
  • siderophores
  • Brucella abortus
  • entF
  • erythritol
  • brucebactin
Date of Defense 2009-05-11
Availability restricted
Abstract
Brucella causes undulant fever in humans and uterine and systemic infection leading to abortions in domestic animals and wild life. For the acquisition of iron in mammalian hosts, species of Brucella are known to produce two siderophores, 2, 3-dihydroxy benzoic acid (2, 3-DHBA) and brucebactin. Inability to synthesize of 2, 3-DHBA affects the ability of pathogen to metabolize erythritol, replicate in trophoblast cells and cause abortion in pregnant ruminant host. The entF gene has been implicated in the unresolved pathway allowing brucebactin biosynthesis in Brucella. The research effort presented in this thesis tries to relate the role of entF in iron acquisition and potential relation with erythritol metabolism by wild type B. abortus 2308. An entF deletion mutant (BAN1) of B. abortus 2308, generated using cre-lox methodology was found to be growth inhibited in iron minimal media compared to wild type strain. Growth inhibition was further enhanced with the addition of an iron chelator or 0.1% erythritol. Compared to wild type strain, no growth inhibition of BAN1 mutant was found in murine J774A.1 macrophages, which suggests that Brucella could acquire iron inside mammalian cells. The entF gene complemented mutant strains of BAN1 (BAN2A and BAN2B) were found to be intermediate in their ability to grow in iron minimal media supplemented with 0.0.05% erythritol, when compared to wild type and BAN1 strain. The results from the present thesis demonstrate that entF gene plays an important role in iron acquisition and erythritol metabolism by B. abortus 2308 under iron limiting conditions.
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