Dry-Milling and Fractionation of Transgenic Maize Seed Tissues with Green Fluorescent Protein as a Tissue Marker

Autor: M. P. Scott, Colin T. Shepherd, Lawrence A. Johnson, Nathalie Vignaux, J. M. Peterson
Rok vydání: 2008
Předmět:
Zdroj: Cereal Chemistry Journal. 85:196-201
ISSN: 0009-0352
DOI: 10.1094/cchem-85-2-0196
Popis: The efficiency of fractionating cereal grains (e.g., dry corn milling) can be evaluated and monitored by quantifying the proportions of seed tissues in each of the recovered fractions. The quantities of individual tissues are typically estimated using indirect methods such as quantifying fiber or ash to indicate pericarp and tip cap contents, and oil to indicate germ content. More direct and reliable methods are possible with tissue-specific markers. We used two transgenic maize lines, one containing the fluorescent protein green fluorescent protein (GFP) variant S65T expressed in endosperm, and the other containing GFP expressed in germ to determine the fate of each tissue in the dry-milling fractionation process. The two lines were dry-milled to produce three fractions (bran-, endosperm-, and germ-rich fractions) and GFP fluorescence was quantified in each fraction to estimate the tissue composition. Using a simplified laboratory dry-milling procedure and our GFP-containing grain, we determined...
Databáze: OpenAIRE