The functionality of α-kafirin promoter and α-kafirin signal peptide
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- 作者：Guoquan Liu ; Kyle C. Lamont ; Norazlina Ahmad…
- 关键词：Endosperm ; specific promoter ; Constitutive promoter ; Signal peptide ; Protein body ; Gene expression ; Sorghum ; Green fluorescent protein
- 刊名：Plant Cell, Tissue and Organ Culture (PCTOC)
- 出版年：2017
- 出版时间：January 2017
- 年：2017
- 卷：128
- 期：1
- 页码：133-143
- 全文大小：
- 刊物类别：Biomedical and Life Sciences
- 刊物主题：Plant Sciences; Plant Physiology; Plant Genetics & Genomics; Plant Pathology;
- 出版者：Springer Netherlands
- ISSN：1573-5044
- 卷排序：128
文摘
Cereal grains offer great potential as a storage system for production of highly valuable proteins using biotechnological approaches, but such applications require tight temporal and spatial control of transgene expression. Towards this aim, we have undertaken a detailed analysis of α-kafirin (α-kaf) promoter and α-kaf signal peptide (sp) in transgenic sorghum plants, using green fluorescent protein gene (gfp) as a reporter. Constructs containing either the α-kaf promoter or the constitutive maize ubiquitin-1 (ubi) promoter driving either gfp or sp-gfp translational fusion were introduced into Sorghum bicolor inbred line Tx430 by particle bombardment. We show for the first time that the α-kaf promoter directs endosperm-specific transgene expression, with activity first detected at 10 days post-anthesis (dpa), peaking at 20 dpa, and remaining active through to physiological maturity. Furthermore, we demonstrate for the first time that the α-kafirin sp is sufficient to direct foreign protein to protein bodies in the endosperm. The evidence is also provided for possible mis-targeting by α-kaf sp in vegetative tissues of transgenic lines with ubi-sp-gfp, resulting in loss of reporter gene translational activity that no GFP signal was observed. These results demonstrate that α-kaf promoter and α-kaf sp are well suited for seed bioengineering to produce recombinant proteins in sorghum endosperm or deposit foreign proteins into sorghum protein bodies.