Determination of phosphinothricin acetyltransferase in genetically transformed canola seed by a two-antibody sandwich enzyme immunoassay |
| |
Authors: | Brigitte Bauer-Weston Arno Schulz Michael M Oelck Raymond J A Deschamps |
| |
Institution: | (1) AgrEvo Canada Inc., 104-108 Research Drive, S7N 3R3 Saskatoon, SK, Canada;(2) Werk Hoechst, Hoechst Schering AgrEvo GmbH, H 872N, D-65926 Frankfurt, Germany;(3) AgrEvo Canada Inc., 295 Henderson Drive, S4N 6C2 Regina, SK, Canada |
| |
Abstract: | We report a quantitative, two-antibody sandwich immunoassay for phosphinothricin acetyltransferase (PAT), the selectable marker
protein. The method yielded a standard curve with a working range of 0 to 5 ng PAT per mL extract. Replicate absorbance values
for standards within a single assay showed a coefficient of variation typically less than 5 percent. Over three separate assays,
the coefficient of variation for the slope and y-intercept of the standard curve was 3.8 and 3.7 percent, respectively. TransgenicBrassica napus L. seed was used to demonstrate the utility of the assay. Non-transgenic seed extracts did not show a positive immunoassay
signal. Determinations of the PAT enzyme conducted on spiked non-transgenic seed extracts repeated in three separate assays
fell in the acceptable range of 80 to 120 percent recovery. Transgenic canola seed, analyzed in three separate assays, showed
a mean PAT enzyme content of 403 ng/g with a standard error of 19 ng/g and a coefficient of variation of 8.1 percent. The
method has also been applied to several other tissues and processed products of canola, maize, and tobacco. |
| |
Keywords: | Brassica napus immunoassay phosphinothricin acetyltransferase transgenic canola |
本文献已被 SpringerLink 等数据库收录! |
|