Use of untranslated region of osmotin gene to enhance transgene expression in plants

Abstract

The present invention provides methods, vectors and gene constructs for enhancing expression of a recombinant nucleic acid sequence in transgenic plants and plant tissues. According to the present invention, nucleic acid sequences are obtained and/or derived from the 5′ and 3′ untranslated regions of genes encoding osmotin proteins and engineered to flank respective portions of a selected coding region of a vector. The vector construct may be introduced into plants and/or plant tissues through conventional procedures, resulting in enhanced expression of the selected coding region. In a preferred embodiment, the selected coding region is a chimeric gene or gene fragment expressing one or more proteins known to impart a level of insecticidal activity to a transgenic plant and/or plant tissue.

1 . A nucleic acid construct comprising at least one structural gene of interest functionally linked to one or more of a UTR selected from the group consisting of SEQ. ID. 1, SEQ. ID. No. 2, SEQ. ID No. 3. and derivatives or functional equivalents of any thereof.  
     
     
         2 . The nucleic acid construct of  claim 1 , wherein the at least one structural gene of interest comprises a gene capable of conferring a non-native phenotype in a plant.  
     
     
         3 . The nucleic acid construct of  claim 1 , wherein the at least one structural gene of interest comprises a gene capable of conferring insecticide or herbicide resistance in a plant.  
     
     
         4 . The nucleic acid construct of  claim 1 , wherein the at least one structural gene of interest comprises an insect resistance gene isolated or derived from the group consisting of SEQ. ID. Nos. 6, 7, 8, 9, and 10.  
     
     
         5 . The construct of  claim 1 , wherein the at least one structural gene of interest comprises SEQ. ID. No. 5 or a derivative thereof.  
     
     
         6 . A plant or plant cell transformed with the nucleic acid construct of  claim 1 .  
     
     
         7 . The plant or plant cell of  claim 6  further comprising an additional structural gene of interest stacked with the at least one gene of interest.  
     
     
         8 . The plant or plant cell of  claim 7 , wherein the additional structural gene of interest is functionally linked to one or more of a UTR selected from the group consisting of SEQ. ID. 1, SEQ. ID. No. 2, SEQ. ID No. 3. and derivatives or functional equivalents of any thereof.  
     
     
         9 . A vector comprising the nucleic acid construct of  claim 1 .  
     
     
         10 . A method for recombinantly producing a peptide or protein comprising: 
 functionally linking a nucleic acid sequence consisting of an osmotin UTR element with a structural gene of interest.    
     
     
         11 . A method of increasing expression of a gene in a plant cell comprising: 
 functionally linking at least one nucleic acid sequence including of an osmotin UTR element comprising at least one of SEQ. ID. Nos. 1-3 with a structural gene of interest to create a nucleic acid construct;    transforming the plant cell with the nucleic acid construct; and    growing the transformed cell under conditions in which the structural gene of interest is expressed.    
     
     
         12 . A recombinant DNA construct comprising a gene of interest and at least of a UTR selected from the group consisting of a 5′ and a 3′ UTR from an osmotin gene.  
     
     
         13 . A 5′ UTR from an osmotin gene or a derivative thereof functionally linked to a heterologous gene.  
     
     
         14 . A 3′ UTR from an osmotin gene or a derivative thereof functionally linked to a heterologous gene.  
     
     
         15 . A recombinant nucleic acid construct comprising 5′ and/or 3′ UTR isolated or derived from an osmotin gene, the 5′ and/or 3′ UTR functionally linked to a structural gene of interest.  
     
     
         16 . The recombinant nucleic acid construct of  claim 15 , wherein the 5′ and/or 3′ UTR is derived from an osmotin gene from tobacco.