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Rearing and Injection of Manduca sexta Larvae to Assess Bacterial Virulence
Published on: December 11, 2012
Stimulation of midgut stem cell proliferation by Manduca sexta alpha-arylphorin
Michael B Blackburn1, Marcia J Loeb, Edward Clark
1Insect Biocontrol Laboratory, Plant Sciences Institute, Agricultural Research Service, USDA, Beltsville, Maryland 20705, USA. blackbm@ba.ars.usda.gov
Abstract:
Extracts of the green-colored perivisceral fat body of newly ecdysed Manduca sexta pupae stimulate mitosis in midgut stem cells of Heliothis virescens cultured in vitro. Using a combination of cation- and anion-exchange chromatography, we have isolated a protein from these fat body extracts that accounts for the observed stem cell proliferation. SDS-PAGE analysis of the protein results in a single band of 77 kDa. Sequences of tryptic peptides from this protein are identical to internal sequences of the storage hexamer alpha-arylphorin. The alpha-arylphorin isolated by our procedure represents a small fraction of the total arylphorin present in the fat body extract. However, it alone seems responsible for the stimulation of mitotic activity in H. virescens midgut stem cells.
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