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Effects of different brush border membrane vesicle isolation protocols on proteomic analysis of Cry1Ac binding proteins from the midgut of Helicoverpa armigera
Author(s) -
Chen LiZhen,
Liang GeMei,
Rector Brian G.,
Zhang Jie,
Wu KongMing,
Guo YuYuan
Publication year - 2008
Publication title -
insect science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.991
H-Index - 45
eISSN - 1744-7917
pISSN - 1672-9609
DOI - 10.1111/j.1744-7917.2008.00238.x
Subject(s) - brush border , cry1ac , helicoverpa armigera , bacillus thuringiensis , biology , midgut , helicoverpa , biochemistry , membrane protein , gel electrophoresis , polyacrylamide gel electrophoresis , vesicle , microbiology and biotechnology , larva , bacteria , membrane , botany , gene , enzyme , genetics , genetically modified crops , transgene
Brush border membrane vesicles (BBMV) isolated from insect midguts have been widely used to study Cry1A binding proteins. Sample preparation is important in two‐dimensional electrophoresis (2‐DE), so to determine a suitable BBMV preparation method in Helicoverpa armigera for 2‐DE, we compared three published BBMV preparation methods mostly used in sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE). All methods yielded similar types and numbers of binding proteins, but in different quantities. The Abdul‐Rauf and Ellar protocol was the best of the three, but had limitations. Sufficient protein quantity is important for research involving limited numbers of insects, such as studies of insect resistance to Bacillus thuringiensis in the field. Consequently, we integrated the three BBMV isolation methods into a single protocol that yielded high quantities of BBMV proteins from H. armigera larval midguts, which proved suitable for 2‐DE analysis.