Extraction and separation of water-soluble proteins from Bacillus thuringiensis-transgenic and non-transgenic maize species by CZE
✍ Scribed by Petra Sázelová; Václav Kašička; Elena Ibáñez; Alejandro Cifuentes
- Publisher
- John Wiley and Sons
- Year
- 2009
- Tongue
- English
- Weight
- 304 KB
- Volume
- 32
- Category
- Article
- ISSN
- 1615-9306
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✦ Synopsis
Abstract
A new method for extraction and CZE separation and characterization of water‐soluble proteins (albumin fraction) from Bacillus thuringiensis‐transgenic and non‐transgenic maize varieties was developed. Protein content in the extracts from maize flours was estimated by UV‐absorbance at 280 nm and Lowry protein assay. Protein extracts were analyzed by CZE in four acidic BGEs (100 mM H~3~PO~4~, 50 mM Tris, pH 2.25; 500 mM acetic acid, pH 2.6; 200 mM formic acid, 200 mM acetic acid, pH 2.05; 200 mM iminodiacetic acid, pH 2.26) using a homemade CZE device equipped with bare fused silica capillary and UV‐absorption detector at 206 nm. Among the tested BGEs, the best resolution of extracted proteins was achieved in isoelectric BGE, 200 mM iminodiacetic acid, pH 2.26. Well‐resolved components of protein extracts were characterized by effective electrophoretic mobilities and corrected (migration times normalized) peak areas. Significant qualitative and quantitative differences in CZE‐UV profiling were found. This approach can potentially be used to distinguish the B. thuringiensis variety from the wild maize in an easy and straightforward way.