Promoter analysis of seed storage protein genes from Canavalia gladiata D.C.
A number of A/T-rich sequences and a CATGCAT/A sequence are contained in the 5'-upstream regions of the genes encoding concanavalin A (Con A) and canavalin, two major seed storage proteins of Canavalia gladiata D.C. To study the role of these sequences in the seed-specific gene expression, we constructed 5'-deletion mutants and examined the transient expression of beta-glucuronidase reporter gene by particle bombardment and the stable expression by Agrobacterium-mediated transformation of tobacco plants. Positive regulatory elements were located in the -894/-602 and -602/-74 regions of the Con A gene, and in the -428/-376, -281/-155 and -155/-50 regions of the canavalin gene. In addition, the results suggested that the A/T-rich sequences in the 5'-upstream region of the Con A gene play a role in transcriptional activation, but that those of the canavalin gene have little effect on the gene expression. The CATGCAT/A sequence was not sufficient by itself for high levels of expression of both the Con A and canavalin genes. The canavalin polypeptide amounted to about 1% of the total extractable protein in the transgenic tobacco seeds, but the Con A polypeptide was not detected in the extractable protein.
Yamamoto
S
S
Department of Biology, Tokyo Metropolitan University, Japan.
Nishihara
M
M
Morikawa
H
H
Yamauchi
D
D
Minamikawa
T
T
D001483:Base Sequence; D003208:Concanavalin A / Q000235:genetics*; D018744:DNA, Plant; D007887:Fabaceae / Q000235:genetics*; D008969:Molecular Sequence Data; D037121:Plant Lectins; D010940:Plant Proteins / Q000235:genetics*; D030821:Plants, Genetically Modified; D010946:Plants, Medicinal; D010947:Plants, Toxic; D011401:Promoter Regions, Genetic; D012639:Seeds / Q000378:metabolism*; D017384:Sequence Deletion; D014026:Nicotiana / Q000235:genetics
D018744:DNA, Plant; D037121:Plant Lectins; D010940:Plant Proteins; C006882:canavalin protein, Canavalia ensiformis; D003208:Concanavalin A