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. 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.
School of Pharmacy, The University of Queensland, Brisbane, QLD 4072, Australia. norazlina@uq.edu.au
Norazlina
N
Ahmad
Rajnesh
R
Sant
Milovan
M
Bokan
Kathryn J
KJ
Steadman
Ian D
ID
Godwin
D018506:Gene Expression Regulation, Plant / Q000502:physiology*; D024861:Genetic Enhancement / Q000379:methods; D010940:Plant Proteins / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000502:physiology*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D021382:Protein Sorting Signals / Q000235:genetics*; D012639:Seeds / Q000502:physiology*; D045868:Sorghum / Q000502:physiology*
D010940:Plant Proteins; D021382:Protein Sorting Signals; C080615:kafirin protein, Sorghum bicolor