|Habibi, Javad - UNIV OF MISSOURI|
|Stuart, Melissa - STILL UNIV|
Submitted to: Congress on In Vitro Biology
Publication Type: Abstract Only
Publication Acceptance Date: April 20, 2005
Publication Date: June 7, 2005
Citation: Habibi, J., Goodman, C.L., Stuart, M.K. 2005. Distribution of elongation factor-1alpha in larval tissues of the fall armyworm, spodoptera frugiperda [abstract]. Congress on In Vitro Biology. p. 34A. Technical Abstract: Elongation factor-1alpha (EF-1a) promotes the delivery of aminoacyl-tRNA to the acceptor site of the ribosome during protein synthesis. It also plays a pivotal role in regulating apoptosis: cultured insect cells that accumulate EF-1a become apoptotic, possibly by enhancing translation of “killer factors” such as caspase enzymes. Mab 7D6, a monoclonal antibody generated to EF-1a from the fall armyworm, inhibits in vitro translation when added to lysates of Sf21 cells. Our long-term goal is to clone a single-chain antibody (scFv) gene encoding Mab 7D6 into the baculovirus to enhance the virus’s potential as a biological control agent. We anticipate that fall armyworm larvae infected by the recombinant viruses will die more quickly than those infected by wild-type viruses. As Mab 7D6 binds to cytoplasmic EF-1a, the antibody may prevent the synthesis of proteins vital to the host cell, or enhance virion production by delaying apoptosis. Because immunologically distinct, tissue-specific forms of EF-1a commonly occur in eukaryotes, tissues of fall armyworm larvae were probed with Mab 7D6 to determine whether the tissues most important for establishing viral infection were recognized. Using western blotting, ELISA, and microscopy techniques, we found that all tissues examined contained measurable amounts of EF-1a reactive with Mab 7D6, although concentrations varied among different cell types within a given tissue. The intensity of the signals was much stronger on the apical part of the columnar epithelial cells, especially on brush-border microvilli, than the basal parts of these cells. No signal was observed on goblet cells and basement membrane.