Serial extraction of endosperm drillings (SEED)—A method for detecting transgenes and proteins in single viable maize kernels

dc.contributor.author Sangtong, Varaporn
dc.contributor.author Mottl, Erik
dc.contributor.author Long, Mary Jane
dc.contributor.author Lee, Michael
dc.contributor.author Scott, M. Paul
dc.contributor.department Agronomy
dc.date 2018-02-18T14:58:58.000
dc.date.accessioned 2020-06-29T23:03:09Z
dc.date.available 2020-06-29T23:03:09Z
dc.date.issued 2001-06-01
dc.description.abstract <p>We have developed a method for detecting a transgene and its protein product in maize endosperm that allows the kernel to be germinated after analysis. This technique could be highly useful for several monocots and dicots. Our method involves first sampling the endosperm with a hand-held rotary grinder so that the embryo is preserved and capable of germination. This tissue is then serially extracted, first with SDS-PAGE sample buffer to extract proteins, then with an aqueous buffer to extract DNA. The product of the transgene can be detected in the first extract by SDS-PAGE with visualization by total protein staining or immuno-blot detection. The second extract can be purified and used as template DNA in PCR reactions to detect the transgene. This method is particularly useful for screening transgenic kernels in breeding experiments and testing for gene silencing in kernels.</p>
dc.description.comments <p>This article is published as Sangtong, Varaporn, Erik C. Mottl, Mary Jane Long, Michael Lee, and M. Paul Scott. "Serial extraction of endosperm drillings (seed)—A method for detecting transgenes and proteins in single viable maize kernels." <em>Plant Molecular Biology Reporter</em> 19, no. 2 (2001): 151-158.</p>
dc.format.mimetype application/pdf
dc.identifier archive/lib.dr.iastate.edu/agron_pubs/176/
dc.identifier.articleid 1168
dc.identifier.contextkey 10438964
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath agron_pubs/176
dc.identifier.uri https://dr.lib.iastate.edu/handle/20.500.12876/4506
dc.language.iso en
dc.source.bitstream archive/lib.dr.iastate.edu/agron_pubs/176/2001_Scott_SerialExtraction.pdf|||Fri Jan 14 21:26:08 UTC 2022
dc.subject.disciplines Agricultural Science
dc.subject.disciplines Agronomy and Crop Sciences
dc.subject.disciplines Plant Breeding and Genetics
dc.subject.keywords ELISA
dc.subject.keywords gene expression
dc.subject.keywords PCR
dc.subject.keywords transgene
dc.subject.keywords Zea mays L.
dc.title Serial extraction of endosperm drillings (SEED)—A method for detecting transgenes and proteins in single viable maize kernels
dc.type article
dc.type.genre article
dspace.entity.type Publication
relation.isAuthorOfPublication 97acee5f-1291-4c27-8929-a8e1617c411d
relation.isOrgUnitOfPublication fdd5c06c-bdbe-469c-a38e-51e664fece7a
File
Original bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
2001_Scott_SerialExtraction.pdf
Size:
605.11 KB
Format:
Adobe Portable Document Format
Description:
Collections