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Identification of midgut microvillar proteins fromTenebrio molitorandSpodoptera frugiperdaby cDNA library screenings with antibodies

Authors
Journal
Journal of Insect Physiology
0022-1910
Publisher
Elsevier
Publication Date
Volume
53
Issue
11
Identifiers
DOI: 10.1016/j.jinsphys.2007.06.007
Keywords
  • Microvillar Membrane
  • Sequencing
  • Antibody Screening
  • Est
  • Coleoptera
  • Lepidoptera
  • Digestion
Disciplines
  • Biology

Abstract

Abstract The objective of this study was to identify midgut microvillar proteins in insects appearing earlier (Coleoptera) and later (Lepidoptera) in evolution. For this, cytoskeleton-free midgut microvillar membrane from Spodoptera frugiperda (Lepidoptera) and Tenebrio molitor (Coleoptera) were used to raise antibodies. These were used for screening midgut cDNA expression libraries. Positive clones were sequenced, assembled and searched for similarities with gene/protein databases. The predicted midgut microvillar proteins from T. molitor were: cockroach allergens (unknown function), peritrophins (peritrophic membrane proteins), digestive enzymes (aminopeptidase, α-mannosidase) and unknown proteins. Predicted S. frugiperda midgut proteins may be grouped into six classes: (a) proteins involved in protection of midgut (thioredoxin peroxidase, aldehyde dehydrogenase, serpin and juvenile hormone epoxide hydrolase); (b) digestive enzymes (astacin, transporter-like amylase, aminopeptidase, and carboxypeptidase); (c) peritrophins; (d) proteins associated with microapocrine secretion (gelsolin, annexin); (e) membrane-tightly bound-cytoskeleton proteins (fimbrin, calmodulin) and (f) unidentified proteins. The novel approach is compared with others and microvillar function is discussed in the light of the predicted proteins.

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