Proteomic Identification of Dengue Virus Binding Proteins in Aedes aegypti Mosquitoes and Aedes albopictus Cells
Affinity chromatography of MG protein extracts. MG proteins were purified from extracts of different Ae. aegypti strains (DMEB, DS3, IBO-11, or Mori) by affinity chromatography using DENV-2, -1, -4 or rE2-DIII-Sepharose 4B column as described in Section 2. Midgut proteins were eluted from DENV-2-Sepharose 4B columns with buffer E containing 1 M NaCl (lines 1-2), or 0.1 M Glycine pH 2.7 (lines 3-4) and from rE2-DIII-Sepharose 4B column with 1 M NaCl (line 5) or 0.1 M Glycine pH 2.7 containing 0.5 M NaCl (line 6-7). Aliquots of 500 μL were collected from each column and proteins were acetone-precipitated and separated by 10% SDS-PAGE and Coomassie Brilliant Blue or silver stained. The apparent molecular weights of these proteins are shown on the right side. Molecular weight markers are shown on the left side.
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