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Expression Pattern of the Alpha-Kafirin Promoter Coupled with a Signal Peptide from Sorghum bicolor L. Moench

Regulatory sequences with endosperm specificity are essential for foreign gene expression in the desired tissue for both grain quality improvement and molecular pharming. In this study, promoters of seed storage α-kafirin genes coupled with signal sequence (ss) were isolated from Sorghum bicolor L....

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Autores principales: Ahmad, Norazlina, Sant, Rajnesh, Bokan, Milovan, Steadman, Kathryn J., Godwin, Ian D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3270457/
https://www.ncbi.nlm.nih.gov/pubmed/22315514
http://dx.doi.org/10.1155/2012/752391
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author Ahmad, Norazlina
Sant, Rajnesh
Bokan, Milovan
Steadman, Kathryn J.
Godwin, Ian D.
author_facet Ahmad, Norazlina
Sant, Rajnesh
Bokan, Milovan
Steadman, Kathryn J.
Godwin, Ian D.
author_sort Ahmad, Norazlina
collection PubMed
description Regulatory sequences with endosperm specificity are essential for foreign gene expression in the desired tissue for both grain quality improvement and molecular pharming. In this study, promoters of seed storage α-kafirin genes coupled with signal sequence (ss) were isolated from Sorghum bicolor L. Moench genomic DNA by PCR. The α-kafirin promoter (α-kaf) contains endosperm specificity-determining motifs, prolamin-box, the O2-box 1, CATC, and TATA boxes required for α-kafirin gene expression in sorghum seeds. The constructs pMB-Ubi-gfp and pMB-kaf-gfp were microprojectile bombarded into various sorghum and sweet corn explants. GFP expression was detected on all explants using the Ubi promoter but only in seeds for the α-kaf promoter. This shows that the α-kaf promoter isolated was functional and demonstrated seed-specific GFP expression. The constructs pMB-Ubi-ss-gfp and pMB-kaf-ss-gfp were also bombarded into the same explants. Detection of GFP expression showed that the signal peptide (SP)::GFP fusion can assemble and fold properly, preserving the fluorescent properties of GFP.
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spelling pubmed-32704572012-02-07 Expression Pattern of the Alpha-Kafirin Promoter Coupled with a Signal Peptide from Sorghum bicolor L. Moench Ahmad, Norazlina Sant, Rajnesh Bokan, Milovan Steadman, Kathryn J. Godwin, Ian D. J Biomed Biotechnol Research Article Regulatory sequences with endosperm specificity are essential for foreign gene expression in the desired tissue for both grain quality improvement and molecular pharming. In this study, promoters of seed storage α-kafirin genes coupled with signal sequence (ss) were isolated from Sorghum bicolor L. Moench genomic DNA by PCR. The α-kafirin promoter (α-kaf) contains endosperm specificity-determining motifs, prolamin-box, the O2-box 1, CATC, and TATA boxes required for α-kafirin gene expression in sorghum seeds. The constructs pMB-Ubi-gfp and pMB-kaf-gfp were microprojectile bombarded into various sorghum and sweet corn explants. GFP expression was detected on all explants using the Ubi promoter but only in seeds for the α-kaf promoter. This shows that the α-kaf promoter isolated was functional and demonstrated seed-specific GFP expression. The constructs pMB-Ubi-ss-gfp and pMB-kaf-ss-gfp were also bombarded into the same explants. Detection of GFP expression showed that the signal peptide (SP)::GFP fusion can assemble and fold properly, preserving the fluorescent properties of GFP. Hindawi Publishing Corporation 2012 2012-01-17 /pmc/articles/PMC3270457/ /pubmed/22315514 http://dx.doi.org/10.1155/2012/752391 Text en Copyright © 2012 Norazlina Ahmad et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Ahmad, Norazlina
Sant, Rajnesh
Bokan, Milovan
Steadman, Kathryn J.
Godwin, Ian D.
Expression Pattern of the Alpha-Kafirin Promoter Coupled with a Signal Peptide from Sorghum bicolor L. Moench
title Expression Pattern of the Alpha-Kafirin Promoter Coupled with a Signal Peptide from Sorghum bicolor L. Moench
title_full Expression Pattern of the Alpha-Kafirin Promoter Coupled with a Signal Peptide from Sorghum bicolor L. Moench
title_fullStr Expression Pattern of the Alpha-Kafirin Promoter Coupled with a Signal Peptide from Sorghum bicolor L. Moench
title_full_unstemmed Expression Pattern of the Alpha-Kafirin Promoter Coupled with a Signal Peptide from Sorghum bicolor L. Moench
title_short Expression Pattern of the Alpha-Kafirin Promoter Coupled with a Signal Peptide from Sorghum bicolor L. Moench
title_sort expression pattern of the alpha-kafirin promoter coupled with a signal peptide from sorghum bicolor l. moench
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3270457/
https://www.ncbi.nlm.nih.gov/pubmed/22315514
http://dx.doi.org/10.1155/2012/752391
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