Abstract
The low level of foreign genes’ expression in transgenic plants is a key factor that limits plant genetic engineering. Because of the critical regulatory activity of the promoters on gene transcription, they are studied extensively to improve the efficiency of the plant transgenic system. The constitutive promoters, such as CaMV 35S promoter, are usually used in plant genetic engineering. But those constitutive promoters continuously express their downstream genes during the whole life span in all the tissues of the host plants. This is not only wasteful to host plant’s energy, but also harmful to host plants and usually affects their agronomic characteristics. In contrast, the seed-specific promoter only expresses its downstream genes from mid to late stage of seed maturation, and there is no expression or much lower expression in other tissues. So the seed-specific promoters are distinguished for their improvement and what they have brought to plant quality engineering. The aim of this article is to characterize a new seed-specific promoter and improve grain quality. The promoter region of β-conglycinin α-subunit gene was isolated from the genomic DNA of soybean Jilin 43 by PCR method, and successfully extended this fragment by TAIL PCR method and obtained the promoter fragment BCSP666. Sequencing analysis showed that the cloned fragment BCSP666 contained all of the motifs, such as RY repeat element, AG/CCCCA motif, TACACAT motif, ACGT motif, A/T rich motif and E-box etc., which constituted the seed-specific promoter activity. Based on this sequencing analysis, the seed-specific promoter activity of the fragment BCSP666 was predicted. And then the seed-specific expression vector pBI121-666, which contained GUS reporter gene, was constructed with the fragment BCSP666. Transformation of Arabidopsis thaliana plants by Agrobacterium-mediated floral-dip method with the recombined vector pBI121-666 was conducted. The transgenic plants were selected on the kanamycin-resistant MS medium, and confirmed by Southern Blot Analysis. Fluorometric and histochemical analysis of GUS enzyme activity of the transgenic Arabidopsis thaliana plants support our original suggestions. Therefore, the seed-specific promoter BCSP666 is obtained and characterized.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Beachy R N, Chen Z L, Horsch R B, Rogers S G, Hoffmann N J, Fraley R T (1985). Accumulation and assembly of soybean β-conglycinin in seeds of transformed petunia plants. The EMBO Journal, 4(12): 3047–3053
Beachy R N, Jarvis N P, Barton K A (1981). Biosynthesis of subunits of the soybean 7S storage protein. J Mol Appl Genet, 1(1): 19–27
Cahoon E B, Carlson T J, Ripp K G, Schweiger B J, Cook G A, Hall S E, Kinney A J (1999). Biosynthetic origin of conjugated double bonds: production of fatty acid components of high-value drying oils in transgenic soybean embryos. Proc Natl Acad Sci USA, 96(22): 12935–12940
Cahoon E B, Marillia E F, Stecca K L, Hall S E, Taylor D C, Kinney A J (2000). Production of fatty acid components of Meadow foam oil in somatic soybean embryos. Plant Physiol, 124: 243–251
Cahoon E B, Ripp K G, Hall S E, Kinney A J (2001). Formation of conjugated Δ8,Δ10-double bonds by Δ12-oleic-acid desaturase-related enzymes. The J Bio Chem, 276(4): 2637–2643
Chamberland S, Daile N, Bernier F (1992). The legumin boxes and the 3′ part of a soybean β-conglycinin promoter are involved in seed gene expression in transgenic tobacco plants. Plant Molecular Biology, 19: 937–949
Chen Z L, Schuler M A, Beachy R N (1986). Functional analysis of regulatory elements in a plant embryo-specific gene. Proc Natl Acad Sci USA, 83: 8560–8564
Clough S J, Bent A F (1998). Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. The Plant Journal, 16(6): 735–743
Hofen R, Willmitzer L (1988). Storage of competent cells for Agrobacterium transformation. Nucleic Acids Research, 16: 9877
Jefferson R A (1987). Assaying chimeric genes in plants: The GUS gene fusion system. Plant Molecular Biology Reporter, 5(4): 387–405
Josefsson L G, Lenman M, Ericson M L, Rask L (1987). Structure of a gene encoding the 1.7S storage protein, Napin from Brassica napus. The Journal of Biological Chemistry, 262: 1196–1208
Kawagoe Y, Murai N (1992). Four distinct nuclear proteins recognize in vitro the proximal promoter of the bean seed storage protein P-phaseolin gene conferring spatial and temporal control. Plant J, 2: 927–936
Liu X Y, Tian S Z, Qin G F, Shen R Y (1997). An improved method for extracting DNA from plants and microorganisms using SDS-CTAB. Journal of Beijing Forestry University, 19(3):101–104 (in Chinese)
Liu Y G, Huang N (1998). Efficient amplification of insert end sequences from Bacterial Artificial Chromosome clones by thermal asymmertric interlaced PCR. Plant Molecular Biology Reporter, 16: 175–181
Liu Y G, Mitsukawa N, Oosumi T, Whittier R F (1995). Efficient isolation and mapping of Arabidopsis thaliana T-DNA insert junction by thermal asymmertric interlaced PCR. The Plant J, 8(3): 457–463
Ochman H, Gerber A S, Hartl D L (1998). Genetic applications of an inverse polymerase chain reaction. Genetics, 120: 621–623
Siebert P D, Chenchik A, Kellogg D E, Lukyanov K A, Lukyanov S A (1995). An improved PCR method for walking in uncloned genomic DNA. Nucleic Acids Research, 23: 1087–1088
Stalberg K, Ellerstrom M, Ezcurra I, Ablov S, Rask L (1996). Disruption of an overlapping E-box/ABRE motif abolished high transcription of the nap A storage protein promoter in transgenic Brassica napus seeds. Planta, 199: 515–519
Vincentz M, Leite A, Neshich G, Vriend G, Mattar C, Barros L, Weinberg D, de Almeida E R, de Carvalho M P, Aragao F, Gander E S (1997). ACGT and vicilin core sequences in a promoter domain required for seed-specific expression of a 2S storage protein gene are recognized by the opaque-2 regulatory protein. Plant Molecular Biology, 34: 879–889
Yoshino M, Kanazawa A, Tsutsumi K I, Nakamura I, Shimamoto Y (2001). Structure and characterization of the gene encoding alpha subunit of soybean beta-conglycinin. Genes Genet Syst, 76(2): 99–105
Author information
Authors and Affiliations
Corresponding author
Additional information
__________
Translated from Acta Agronomoca Sinica, 2005, 31(1): 11–17 [译自: 作物学报]
About this article
Cite this article
Caiyin, Q., Li, M., Wei, D. et al. Isolation and sequencing analysis on the seed-specific promoter from soybean. Front. Agric. China 1, 17–23 (2007). https://doi.org/10.1007/s11703-007-0003-1
Issue Date:
DOI: https://doi.org/10.1007/s11703-007-0003-1