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KMID : 0578319930030020165
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Molecules and Cells
1993 Volume.3 No. 2 p.165 ~ p.170
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Interaction of Nuclear Factor with the 5´ Upstream Region of Soybean Glycinin Gene
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Kim, Chung Ho
Kang, Hong-Gu/Choi, Yang Do
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Abstract
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To study the regulatory mechanism of soybean glycinin expression genes gemonic DNA clone of Gy2 was isolated -by plaque hybridization of a genomic library with the glycinin cDNA probe. Nucleotide sequence of the 5´ upstream region of the Gy2 gene revealed some of the putative regulatory elements such as storage protein-specific elements, embryo factor-binding site, conglycinin enhancer, and OPAQUE-2 binding site in addition to the canonical promoter elements. Gel retardation assay with the nuclear extract prepared from developing seeds demonstrated that nuclear proteins bind to DNA fragments up to -220 by from the transcription initiation site containing those putative regulatory elements. The on-radioactive probes compete off the nuclear factor bound to the radioactive probes, demonstrating their sequence-specific interaction. This region could be separated at -85 into two domains which do not compete each other for nuclear protein binding. This suggests that they interact with different nuclear proteins. DNase I foot print analysis showed that the region around -61 was protected from DNase I digestion, which contains a OPAQUE-2 binding sequence. These results suggested the presence of regulatory cis-acting sequences for trans-acting protein factors for regulation of Gy2 expression. Combination of these cis- and trans-acting factors could maintain tissue-and development-specific gene expression pattern of the glycinin gene.
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KEYWORD
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