Plant embryo—and aleurone—specific promoter

Details Plant embryo—and aleurone—specific promoter Plant embryo—and aleurone—specific promoter

1999-10-13

2001-10-23

McGarry, Sean (Department: 1635)

Chemistry: molecular biology and microbiology

Plant cell or cell line, per se ; composition thereof;...

Plant cell or cell line, per se, contains exogenous or...

C435S320100, C435S410000, C536S023100, C536S023600, C536S024100

Reexamination Certificate

active

06306656

ABSTRACT:

BACKGROUND OF THE INVENTION
Supplementation of hydrolytic enzymes in farm animal feed can improve agricultural efficiency by decreasing the amount of fecal material, and therefore the waste removal cost and environmental hazard. However, the high cost of producing such enzymes precludes its low value-added use as a feed supplement. Since rice bran (including the rice embryo and aleurone) is a major component of many animal feeds, expression of hydrolytic enzymes in rice tissues may provide a cost-effective means of decreasing the environmental impact of animal farming.
SUMMARY OF THE INVENTION
The invention is based on the discovery of a new barley promoter (as part of the gene designated aba45) that can promote transcription in rice embryo cells. Thus, the promoter of the invention can be used to express hydrolytic enzymes in rice bran tissues.
Accordingly, the invention features an isolated nucleic acid comprising SEQ ID NO:1, a barley aba45 promoter sequence that can promote expression of a heterologous gene in rice cells. The full sequence of the aba45 gene of the barley
Hordeum vulgare
cv. Himalaya is shown in FIG.
1
.
The invention also features an isolated nucleic acid having a sequence that hybridizes to SEQ ID NO:2 under stringent conditions or that is at least 70% (e.g., at least 80, 90, 95, or 99%) identical to SEQ ID NO:2, the sequence promoting transcription in a plant cell (i.e., is capable of promoting transcription in a plant cell when operably linked to an open reading frame). SEQ ID NO:2 is the portion of the promoter upstream of the cDNA start site (see FIG.
1
and the description thereof below). The plant cell can be a plant embryo cell (i.e., derived from a plant embryo) or a plant aleurone cell (i.e., derived from a plant aleurone).
The invention also includes any vectors or transformed cells which contain a nucleic acid of the invention. Vectors include nucleic acid vectors, such as expression plasmids, or viral vectors. Transformed cells include eukaryotic and prokaryotic cells.
By “hybridizes under stringent conditions” is meant specific and non-covalent binding to an immobilized reference nucleic acids in the presence of 0.2×SSC (1.75 g/1 NaCl, 0.88 g/1 Na
3
citrate. 2H
2
O; pH 7.0) and 0.1% (w/v) sodium dodecylsulfate at 68° C.
An isolated nucleic acid is one which is separated from other nucleic acid molecules which are present in the natural source of the nucleic acid. An isolated nucleic acid is free of sequences which naturally flank the nucleic acid (i.e., sequences flanking the 5′ and 3′ ends of the nucleic acid) in the genomic DNA of the organism from which the nucleic acid is derived.
Where a nucleic acid molecule is said to have a specific percent identity to a reference nucleic acid molecule, the percent identity is determined by the algorithm of Myers and Miller, CABIOS, 1989, which is embodied in the ALIGN program (version 2.0), or its equivalent, using a gap length penalty of 12 and a gap penalty of 4 where such parameters are required. All other parameters are set to their default positions. Access to ALIGN is readily available. See, e.g., http://www2.igh.cnrs.fr\, bin/align-guess.cgi on the Internet.
The promoter sequence of the invention can be introduced into a variety of plant expression vectors for expressing exogenous proteins in plant embryo or aleurone tissue. Such exogenous proteins include hydrolytic enzymes such as phytases, cellulases, xylanases, &bgr;-glucanases, amylases, lipases, proteases, or any other polypeptide that increases the commercial value of the plant as an animal feed component. More specifically, the new promoter sequence and an open reading frame to which it is operably linked can be integrated into a plant genome (e.g., a rice genome) to produce transgenic plants that express hydrolytic enzymes. Methods of delivering nucleic acids into a plant cell, whether for transient or stable expression, are well known in the art of plant biology.
Other features or advantages of the invention will be apparent from the following detailed description, the drawing, and also from the claims.


REFERENCES:
Jin-Hao Liu et al., Identification and Characterization of a Novel Barley Gene that is ABA-Inducible and Expressed Specifically in embryo and Aleurone, May 1999, Journal of Experimental Botany, vol. 50, No. 334, pp. 727-728.*
Hill, Robert D. et al., “Abscisic Acid Structure-Activity Relationships in Barley Aleurone Layers and Protoplasts,” Plant Physiology, vol. 108, No. 2, pp. 573-579, 1995.
Liu, Jin-Hao et al., “Post-transcriptional regulation of bifunctional &agr;-amylase/subtilisin inhibitor expression in barley embryos by abscisic acid,” Plant Molecular Biology, vol. 29, No. 5, pp. 1087-1091, 1995.
Luo, Ma et al., “Characterization of a Gene Family Encoding Abscisic Acid—and Environmental Stress-inducible Proteins of Alfalfa,” The Journal of Biological Chemistry, vol. 267, No. 22, pp. 15367-15374, 1992.
Shen, Qingxi et al., “Functional Dissection of an Abscisic Acid (ABA)—Inducible Gene Reveals Two Independent ABA-Responsive Complexes Each Containing a G-Box . . . ,” The Plant Cell, vol. 7, No. 3, pp. 295-307, 1995.
Skriver, Karen et al., “Cis-acting DNA elements responsive to gibberellin and its antagonist abscisic acid,” Proc. Natl. Acad. Sci. U.S.A. , vol. 88, No. 16, pp. 7266-7270, 1991.

Affiliated with

Also associated with

Rating

Plant embryo—and aleurone—specific promoter does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Plant embryo—and aleurone—specific promoter, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Plant embryo—and aleurone—specific promoter will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-2602108