Development of a kit for diagnosing AtCYP78A7 protein in abiotic-tolerant transgenic rice overexpressing AtCYP78A7

Cited 0 time in scopus
Metadata Downloads
Title
Development of a kit for diagnosing AtCYP78A7 protein in abiotic-tolerant transgenic rice overexpressing AtCYP78A7
Author(s)
K H Nam; Jung Ho ParkIn Soon Pack; H B Kim; Chang-Gi Kim
Bibliographic Citation
Journal of Life Science, vol. 28, no. 7, pp. 835-840
Publication Year
2018
Abstract
Quantitative determination of the protein expression levels is one of the most important parts in assessment of the safety of foods derived from genetically modified (GM) crops. Overexpression of AtCYP78A7, a gene encoding cytochrome P450 protein, has been reported to improve tolerance to abiotic stress, such as drought and salt stress, in transgenic rice (Oryza sativa L.). In the present study, an enzyme-linked immunosorbent assay (ELISA) kit for diagnosing AtCYP78A7 protein including AtCYP78A7-specific monoclonal antibody was developed. GST-AtCYP78A7 recombinant protein was induced and purified by affinity column. Four monoclonal antibodies (mAb 6A7, mAb 4C2, mAb 11H6, and mAb 7E8) against recombinant protein were also produced and biotinylated with avidin-HRP. After pairing test using GST-AtCYP78A7 protein and lysate of rice samples, mAb 4C2 and mAb 7E8 were selected as a capture antibody and a detecting antibody, respectively, for ELISA kit. Product test using rice samples indicated that percentages of detected protein in total protein were greater than 0.1% in AtCYP78A7-overexpressing transgenic rice (Line 10B-5 and 18A-4), whereas those in negative control non-transgenic rice (Ilpum and Hwayoung) were less than 0.1%. The ELISA kit developed in this study can be useful for the rapid detection and safety assessment of transgenic rice overexpressing AtCYP78A7.
Keyword
ELISAgenetically modifided cropmonoclonal antibodysafety assessment
ISSN
I000-0128
Publisher
South Korea
DOI
http://dx.doi.org/10.5352/JLS.2018.28.7.835
Type
Article
Appears in Collections:
Ochang Branch Institute > Division of Bioinfrastructure > Bio-Evaluation Center > 1. Journal Articles
Files in This Item:
  • There are no files associated with this item.


Items in OpenAccess@KRIBB are protected by copyright, with all rights reserved, unless otherwise indicated.