Plant promoter derived from luminal binding protein gene and methods for its use

Abstract

The invention provides a luminal binding protein promoter (PmBiPPro1), deletions thereof, and variants thereof. The promoter is useful for, among other things, directing the expression of transgenes.

1 . A recombinant promoter, comprising a nucleic acid sequence selected from the group consisting of: 
 (a) a nucleic acid sequences selected from the group consisting of SEQ ID NOS: 16,17, 18; and 31;    (b) a nucleic acid sequence that shares at least 50% sequence identity with the nucleic acid sequences shown in (a); and    (c) a nucleic acid sequence that comprises at least 20 consecutive nucleic acid residues of the nucleic acid sequences shown in (a), wherein the promoter is capable of driving the expression of an ORF that is operably linked to the promoter.    
     
     
         2 . The promoter of  claim 1 , comprising the nucleic acid sequence shown in SEQ ID NO: 16.  
     
     
         3 . The promoter of  claim 1 , comprising the nucleic acid sequence shown in SEQ ID NO: 17.  
     
     
         4 . The promoter of  claim 1 , comprising the nucleic acid sequence shown in SEQ ID NO: 18.  
     
     
         5 . The promoter of  claim 1 , comprising the nucleic acid sequence shown in SEQ ID NO: 31.  
     
     
         6 . A vector, comprising a promoter as recited in  claim 1 .  
     
     
         7 . A host cell, comprising a vector as recited in  claim 6 .  
     
     
         8 . A transgenic plant, comprising a host cell as recited in  claim 7 .  
     
     
         9 . A transgene, comprising the promoter of  claim 1  and at least one ORF operably linked to the promoter.  
     
     
         10 . A vector, comprising a transgene as recited in  claim 9 .  
     
     
         11 . A plant cell, comprising a vector as recited in  claim 10 .  
     
     
         12 . A transgene according to  claim 9 , wherein the ORF encodes a cationic peptide.  
     
     
         13 . The plant cell of  claim 11 , wherein the plant cell is obtained from a plant selected from the group consisting of: maize, wheat, rice, millet, tobacco, sorghum, rye, barley, brassica, sunflower, seaweeds, lemna, oat, soybean, cotton, legumes, rape/canola, alfalfa, flax, sunflower, safflower, brassica, cotton, flax, peanut, and clover; lettuce, tomato, cucurbits, cassava, potato, carrot, radish, pea, lentil, cabbage, cauliflower, broccoli, Brussel sprouts, peppers and other vegetables; citrus, apples, pears, peaches, apricots, walnuts, and other fruit trees; orchids, carnations, roses, and other flowers; cacao; poplar, elms, and other deciduous trees; pine, Douglas-fir, spruce, and other conifers; turf grasses; cacao; and rubber trees and other members of the genus Hevea.  
     
     
         14 . A method for expressing at least one protein in a host cell, comprising: 
 provide a recombinant promoter as recited in  claim 1;     operably linking an ORF to the promoter to produce a transgene;    introducing the transgene into a vector; and    introducing the vector into a host cell, wherein the host cell produces a protein from the ORF.    
     
     
         15 . The method according to  claim 14 , wherein the host cell is a plant host cell.  
     
     
         16 . A protein produced according to the method of  claim 14 .  
     
     
         17 . The protein according to  claim 14 , wherein the protein is a cationic peptide.  
     
     
         18 . A promoter according to  claim 1 , wherein the promoter is inducible.  
     
     
         19 . The promoter of  claim 1 , wherein the promoter is inducible at low temperatures.  
     
     
         20 . The promoter of  claim 1 , wherein the promoter is developmentally specific.  
     
     
         21 . The promoter of  claim 20 , wherein the promoter is expressed in actively dividing cells.  
     
     
         22 . The promoter of  claim 1 , wherein the promoter is wound-inducible.  
     
     
         23 . An isolated promoter, comprising at least 8 promoter elements selected from the group consisting of E-box motifs (SEQ ID NO: 1), MNF1 elements (SEQ ID NO: 28), POLLEN1LELAT52 elements (SEQ ID NO: 29) ROOTMOTIF elements (SEQ ID NO: 30), 2SSEEDPROTBANAP elements (SEQ ID NO: 32), BOXIIPCCHS elements (SEQ ID NO: 33), ASF1MOTIF elements (SEQ ID NO: 34), ACGT-core elements (SEQ ID NO: 4), CAAT-boxes (SEQ ID NO: 9), CANABNNAPA elements (SEQ ID NO: 12), HEXMOTIF elements (SEQ ID NO: 27), and duplicates thereof, wherein the promoter displays promoter activity.  
     
     
         24 . The isolated promoter of  claim 23 , wherein the promoter comprises at least 10 promoter elements.  
     
     
         25 . The isolated promoter of  claim 23 , comprising promoter elements in the following order: 3′-ACGT-core element (SEQ ID NO: 4), E-box motif (SEQ ID NO: 1), CAAT-box (SEQ ID NO: 9), 2SSEEDPROTBANAP element(SEQ ID NO: 32), CANABNNAPA element (SEQ ID NO: 12), HEXMOTIF element (SEQ ID NO: 27), CAAT-box (SEQ ID NO: 9), BOXIIPCCHS element (SEQ ID NO: 33), E-box motif (SEQ ID NO: 1), ASF1MOTIF element (SEQ ID NO: 34), POLLEN1LELAT52 element (SEQ ID NO: 29), and MNF1 element (SEQ ID NO: 28)-5′.  
     
     
         26 . A transgene, comprising the promoter of  claim 25  operably linked to an ORF.  
     
     
         27 . A host cell, comprising the transgene of  claim 26 .  
     
     
         28 . A transgenic plant, comprising the host cell of  claim 27 .  
     
     
         29 . The transgenic plant of  claim 28 , wherein the plant is selected from the group consisting of maize, wheat, rice, millet, tobacco, sorghum, rye, barley, brassica, sunflower, seaweeds, lemna, oat, soybean, cotton, legumes, rape/canola, alfalfa, flax, sunflower, safflower, brassica, cotton, flax, peanut, and clover; lettuce, tomato, cucurbits, cassava, potato, carrot, radish, pea, lentil, cabbage, cauliflower, broccoli, Brussel sprouts, peppers and other vegetables; citrus, apples, pears, peaches, apricots, walnuts, and other fruit trees; orchids, carnations, roses, and other flowers; cacao; poplar, elms, and other deciduous trees; pine, Douglas-fir, spruce, and other conifers; turf grasses; cacao; and rubber trees and other members of the genus Hevea.  
     
     
         30 . The transgene according to  claim 26 , wherein the ORF encodes a cationic peptide.  
     
     
         31 . The transgene according to  claim 26 , wherein the ORF encodes the proteins shown in SEQ ID NO: 36.  
     
     
         32 . A method for expressing a protein in a plant, the method comprising: 
 providing a transgene plant according to claim  8 ; and    growing the plant under conditions in which to the plant produces the protein.    
     
     
         33 . The method of  claim 32 , further comprising wounding the plant.  
     
     
         34 . The method of  claim 32 , further comprising growing the plant at a cold temperature.  
     
     
         35 . A method for expressing a protein in a plant, the method comprising: 
 growing a plant according to claim  28 ; wherein the plant produces the protein.    
     
     
         36 . The method of  claim 35 , further comprising wounding the plant.  
     
     
         37 . The method of  claim 35 , further comprising growing the plant in cold temperatures.  
     
     
         38 . The method of  claim 36 , further comprising wounding a plant part and placing the plant part in a cold temperature.