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Generation and molecular analysis of dominant negative alleles of anthrax lethal factor in Drosophila


Anthrax, a disease caused by the bacterium Bacillus Anthracis, poses a threat worldwide due to its debilitating symptoms and lethality. The toxins released by the bacteria (PA, LF, EF) enter cells, cause most symptoms and allow the bacterial growth. LF is a metalloprotease, which cleaves and inactivates MKK in humans, therefore inhibiting MAPK signaling in many cell types. EF is a potent adenylate cyclase and PA facilitates the entry of LF and EF into host cells. In addition to the classic antibiotic and vaccine therapies, one strategy to combat anthrax would be to introduce dominant negative toxins, which could antagonize the wild type toxins, and potentially save lives in the later stages of the disease when antibiotics can no longer prevent death. Model systems such as Drosophila Melanogaster, can be used to study the activity of anthrax and other bacterial toxins, and provide a way to screen such Dominant-negative alleles. We used the Novel OVerexpression Allele screening method to generate several LF alleles which exhibit dominant negative activity as revealed by wing phenotypes in flies. We then attempted to analyze the molecular lesion causing the novel activity of these mutants

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