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Nucleic Acids Research, 1990, Vol. 18, No. 23 6845-6852
© 1990

Articles

Multiple protein factors bind to a rice glutelin promoter region

Soo Young Kim and Ray Wu*

Section of Biochemistry, Molecular and Cell Biology, Cornell University UMR 49 CNRS-ENS, 46 Allèe d'ltalie, 69364 Lyon Cedex 07, France

* To whom correspondence should be addressed

Received September 14, 1990. Revised October 31, 1990. Accepted October 31, 1990.

Our goal is to identify cis-acting elements in the regulatory region of the major seed storage protein gene in rice. A glutelin gene (pGL5-1) has been cloned by screening a rice genomlc DNA library with synthetic ollgonucleotides and with an amplified DNA fragment. A transient expression assay using immature rice seeds shows that its 5' flanking sequence can direct the synthesis of ß-glucuronidase (GUS) when fused upstream of the GUS coding region. Gel-retardation assays were performed to study protein-DNA interactions between putative regulatory sequences of pGL5-1 and nuclear proteins from Immature rice seeds. We demonstrate that at least six protein-DNA complexes are formed between the 5' flanking sequence of pGL5-1 (–677 to –45) and nuclear protein factors. By subsequent DNase I-footprintlng analyses we defined several protein-binding regions. Two of the protein-binding sequences contain the TGAGTCA motif, which is also present in the –300 element found in the 5' flanking sequences of several storage protein genes of other crop plants, and to which the transcription factors jun and GCN4 bind.


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