Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or... – The polynucleotide contains a tissue – organ – or cell...
Reexamination Certificate
2000-03-17
2003-11-04
Bui, Phuong T. (Department: 1638)
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
The polynucleotide contains a tissue, organ, or cell...
C800S278000, C800S298000, C800S309000, C435S419000, C435S468000, C435S320100, C536S023600, C536S024100
Reexamination Certificate
active
06642438
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to novel banana fruit-associated promoters, a melon actin promoter and a banana fruit-associated/melon actin fusion promoter. The invention also relates to heterologous nucleic acid constructs, vectors, kits, and transformation methods employing such promoters. The invention further relates to transgenic plant cells and plants transformed with heterologous nucleic acid constructs comprising the promoters and methods for screening plant promoters in various types of plant tissue using a transient expression assay.
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BACKGROUND OF THE INVENTION
Transcriptional regulatory sequences or promoters that regulate gene expression in plants are essential elements of plant genetic engineering. Several examples of promoters useful for the expression of heterologous genes in plants are now available (Zhu, et al., 1995; Ni, et al., 1995).
Most promoters are from about 500-1500 bases. Promoters for expressing a heterologous gene sequence in plants can be derived from plant DNA, e.g., the cauliflower heat shock protein 80 (hsp80, Brunke and Wilson, 1993; U.S. Pat. No. 5,612,472), or from other sources, for example, plant viruses e.g., the 35S cauliflower mosaic virus promoter, or bacteria which infect plants, e.g., the nopaline synthase (nos) promoter (Rogers, 1991), the octopine synthase (ocs) promoter (Leisner and Gelvin, 1988) and the mannopine synthase (mas) promoter from Agrobacterium.
Expression of heterologous genes or selected sequences of genes in transgenic plants has typically involved the use of constitutive promoters, which drive the expression of a product throughout the plant at all times and in most tissues (e.g., hsp80), the tomato ubiquitin promoter (Picton, et al., 1993), and the raspberry E4 promoter (U.S. Pat. Nos. 5,783,393; and 5,783,394).
A limited number of inducible and/or tissue specific promoters are known. Promoters that provide fruit-specific expression include the E4 and E8 promoter from tomato (Cordes, et al., 1989; Bestwick, et al., 1995; U.S. Pat. No. 5, 859,330). Another fruit-specific promoter is the tomato 2A11 gene promoter. It has been demonstrated that nucleic acid sequences placed under the regulatory control of the 5′ non-coding region of the tomato 2A11 gene (Van Haaren, 1993) are preferentially transcribed in developing fruit tissue. Fruit specific regulation of the kiwifruit actinidin promoter has been reported to be conserved in transgenic petunia plants (Lin, et al., 1993).
Pectate lyase (PEL) has been previously identified as fruit- and ripening-associated in banana (Dominguez-Puigjaner et al., 1997; Medina-Suarez et al., 1997), and has recently been associated with breakdown of cell wall components and subsequent fruit softening during strawberry fruit ripening (Medina-Escobar et al., 1997).
Ethylene is a plant hormone influencing many aspects of plant growth and development, and is known to play a major role in the ripening process in fruits and vegetables. A large amount of ethylene is also produced following trauma caused by chemicals, temperature extremes, water stress, ultraviolet light, insect damage, disease, or mechanical wounding. In some tissues, exposure to only a small amount of ethylene may cause an avalanche of ethylene production in adjacent plants or plant tissues such as fresh produce. This autocatalytic effect can be very pronounced and lead to loss of fruit quality during transportation and storage.
In plants, methionine is converted to AdoMet, which is converted to ACC, which is converted to ethylene. AdoMet is synthesized via a condensation reaction between methionine and Adenosine triphosphate (ATP). A bacterial enzyme, AdoMet hydrolase (AdoMetase), which is normally not present in plant tissue, hydrolyzes AdoMet to homoserine and MTA, both of which are recycled to methionine. Plant transformation vectors, tomato fruit-specific promoters and methods of transforming plants with heterologous nucleic acid constructs effective to express AdoMetase (also termed “SAMase”) in plant cells and thereby modulate ethylene expression, have been described. See, e.g. co-owned U.S. Pat. Nos. 5,416,250; 5,589,623; 5,723,746; 5,750,864; and 5,859,330, expressly incorporated by reference, herein.
A need exists for constitutive promoters of plant origin and for plant promoters that are functional in fruit, and are capable of providing high level expression of heterologous genes in the cells of fruit.
SUMMARY OF THE INVENTION
Applicants have identified novel banana fruit-associated promoters designated in the present application as “TRX” and “PEL”, a melon actin promoter, designated “mACTIN” and me
Clendennen Stephanie K.
Kellogg Jill A.
Mathews Helena V.
Phan Chau B.
Webb Nancy M.
Brunelle Jan P.
Bui Phuong T.
Exelixis Plant Sciences, Inc.
Ibrahim Medina A.
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