Construction and evaluation of a maize (Zea mays) chimaeric promoter with activity in kernel endosperm and embryo

dc.contributor.author Shepherd, Colin
dc.contributor.author Scott, M. Paul
dc.contributor.department Agronomy
dc.date 2018-02-18T14:50:08.000
dc.date.accessioned 2020-06-29T23:02:54Z
dc.date.available 2020-06-29T23:02:54Z
dc.date.issued 2009-03-01
dc.description.abstract <p>Chimaeric promoters contain DNA sequences from different promoters. Chimaeric promoters are developed to increase the level of recombinant protein expression, to precisely control transgene activity or to combat homology-based gene silencing. Sets of chimaeric promoters, each containing different lengths of DNA from maize (<em>Zea mays</em>) 27zn (27 kDa γ-zein) endosperm-specific promoter and the Glb1 (Globulin-1) embryo-specific promoter were created and tested in a transient expression assay of GFP (green fluorescent protein). Promoter fragments with the highest activity were combined to create the chimaeric promoter A27znGlb1. In the context of the chimaeric promoter, the selected Glb1 promoter fragment was necessary and sufficient to activate expression in embryo tissue and was functionally equivalent to the native Glb1 promoter. Similarly, the selected 27zn promoter fragment in the chimaeric promoter was necessary and sufficient to activate expression in endosperm tissue and was functionally equivalent to the native 27zn promoter. Maize transgenic plants containing the A27znGlb1 chimaeric promoter fused to GFP were produced to characterize this promoter <em>in vivo</em>. Quantitative reverse-transcriptase PCR was used to determine that the promoter was active in the embryo, endosperm, pericarp and immature leaf tissues. GFP activity in plants containing the chimaeric promoter was not significantly different in endosperm than the activity of GFP fused to the full-length 27zn promoter, nor was it different in embryo from the activity of GFP fused to the full-length Glb1 promoter. Transgene copy numbers were shown to be between 4 and 12 copies in different events.</p>
dc.description.comments <p>This article is published as Shepherd, Colin T. and Scott, M. Paul. (2009), Construction and evaluation of a maize (<em>Zea mays</em>) chimaeric promoter with activity in kernel endosperm and embryo. <em>Biotechnology and Applied Biochemistry</em>, 52: 233–243, doi:<a href="http://dx.doi.org/10.1042/BA20070269" target="_blank">10.1042/BA20070269</a>.</p>
dc.format.mimetype application/pdf
dc.identifier archive/lib.dr.iastate.edu/agron_pubs/144/
dc.identifier.articleid 1159
dc.identifier.contextkey 10425725
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath agron_pubs/144
dc.identifier.uri https://dr.lib.iastate.edu/handle/20.500.12876/4471
dc.language.iso en
dc.source.bitstream archive/lib.dr.iastate.edu/agron_pubs/144/2009_Scott_ConstructionEvaluation.pdf|||Fri Jan 14 20:19:40 UTC 2022
dc.source.uri 10.1042/BA20070269
dc.subject.disciplines Agricultural Science
dc.subject.disciplines Agronomy and Crop Sciences
dc.subject.disciplines Genetics and Genomics
dc.subject.disciplines Plant Breeding and Genetics
dc.subject.keywords chimaeric (27 kDa γ-zein+Globulin-1) promoter
dc.subject.keywords kernel embryo
dc.subject.keywords kernel endosperm
dc.subject.keywords maize (Zea mays)
dc.subject.keywords recombinant protein expression
dc.subject.keywords transgene
dc.title Construction and evaluation of a maize (Zea mays) chimaeric promoter with activity in kernel endosperm and embryo
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
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