Evaluation of an ELISA assay for rapid detection and quantification of neomycin phosphotransferase II in transgenic plants

Nagel R.J., Manners J.M. and Birch R.G. (1992) Evaluation of an ELISA assay for rapid detection and quantification of neomycin phosphotransferase II in transgenic plants. Plant Molecular Biology Reporter, 10 3: 263-272. doi:10.1007/BF02668359


Author Nagel R.J.
Manners J.M.
Birch R.G.
Title Evaluation of an ELISA assay for rapid detection and quantification of neomycin phosphotransferase II in transgenic plants
Journal name Plant Molecular Biology Reporter   Check publisher's open access policy
ISSN 0735-9640
Publication date 1992-01-01
Sub-type Article (original research)
DOI 10.1007/BF02668359
Open Access Status Not yet assessed
Volume 10
Issue 3
Start page 263
End page 272
Total pages 10
Publisher Kluwer Academic Publishers
Subject 1303 Specialist Studies in Education
1300 Biochemistry, Genetics and Molecular Biology
1110 Nursing
1312 Molecular Biology
Abstract Neomycin phosphotransferase II (neo) is a selectable marker gene used extensively in plant transformation experiments. Here we evaluate immunological detection of its gene product (NPTII) as an alternative to widely used radioactive assays. We have taken a commercially available non-radioactive NPTII Enzyme linked-Immunosorbant Assay (ELISA) kit, modified the protocol for application to plant tissues, and used it to quantify levels of NPTII protein in transformed plants. The ELISA proved safe, economical and convenient to reliably screen and quantify NPTII protein in large numbers of plant samples. The sensitivity of the ELISA for NPTII detection in tobacco plants is at least an order of magnitude greater than a widely used radioactive gel assay. Using three replicates per sample, standard errors are low and the assay is highly reproducibleover time for tissue-cultured tobacco. However, background readings varied with plant species, and also with plant age for untransformed glasshouse-grown tobacco. It is therefore essential to ensure that untransformed controls are closely matched to test plant.
Keyword ELISA
gene expression
neo reporter gene
Nicotiana tabacum
NPT II assay
Saccharum spp
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collection: Scopus Import - Archived
 
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