Nucleic acids encoding chimeric polypeptides allowing expression of plant-noxious proteins

Abstract

This invention relates to nucleic acids encoding chimeric polypeptides comprising vacuole targeting sequences and plant-noxious sequences and especially pest control proteins. The polypeptides are useful in methods for targeting non-vacuolar harmful proteins to plant vacuoles. Chimeric polypeptides of the invention containing pest control proteins are useful for conferring pest resistance on plants and in the production of compositions useful as pesticides.

1 - 64 . (canceled)  
     
     
         65 . An isolated nucleic acid molecule comprising a polynucleotide sequence encoding a vacuole targeting polypeptide operably linked to a polynucleotide sequence encoding a pest control polypeptide.  
     
     
         66 . The nucleic acid molecule of  claim 65 , wherein the vacuole targeting polypeptide is an N-terminal targeting polypeptide.  
     
     
         67 . The nucleic acid molecule of  claim 65 , wherein the vacuole targeting polypeptide is a potato proteinase inhibitor polypeptide.  
     
     
         68 . The nucleic acid molecule of  claim 65 , wherein the pest control polypeptide is a biotin binding protein.  
     
     
         69 . The nucleic acid molecule of  claim 68 , wherein the biotin binding protein is a functional variant of streptavidin.  
     
     
         70 . The nucleic acid molecule of  claim 69 , wherein the functional variant of streptavidin associates with biotin to form a complex with a dissociation constant of 10 −6  M.  
     
     
         71 . The nucleic acid molecule of  claim 69 , wherein the functional variant of streptavidin has 80% homology to a sequence selected from the group consisting of SEQ ID NOS: 3 and 10.  
     
     
         72 . The nucleic acid molecule of  claim 69 , wherein the functional variant of streptavidin has 95% homology to a sequence selected from the group consisting of SEQ ID NOS: 3 and 10.  
     
     
         73 . The nucleic acid molecule of  claim 68 , wherein the biotin binding protein is a functional variant of avidin.  
     
     
         74 . The nucleic acid molecule of  claim 73 , wherein the functional variant of avidin associates with biotin form a complex with a dissociation constant of 10 −6  M.  
     
     
         75 . The nucleic acid molecule of  claim 73 , wherein the functional variant of streptavidin has 80% homology to SEQ ID NO: 2.  
     
     
         76 . The nucleic acid molecule of  claim 73 , wherein the functional variant of streptavidin has 95% homology to SEQ ID NO: 2.  
     
     
         77 . An expression vector comprising the nucleic acid molecule according to  claim 65 .  
     
     
         78 . A host cell transformed with the vector according to  claim 77 .  
     
     
         79 . The host cell according to  claim 78 , wherein said cell is a plant cell.  
     
     
         80 . A transgenic plant comprising the nucleic acid molecule according to  claim 65 .  
     
     
         81 . Seed that is the product of the plant according to  claim 80 .  
     
     
         82 . A method for producing a vacuole targeting polypeptide operably linked to a pest control polypeptide, said method comprising the steps of: 
 (a) culturing a host cell which has been transformed with a vector comprising an isolated nucleic acid molecule comprising a polynucleotide sequence encoding a vacuole targeting polypeptide operably linked to a polynucleotide sequence encoding a pest control polypeptide; and    (b) recovering the expressed vacuole targeting polypeptide operably linked to a pest control polypeptide.    
     
     
         83 . A method for producing a pest resistant plant, said method comprising transforming the plant genome to include at least one nucleic acid molecule comprising a polynucleotide sequence encoding a vacuole targeting polypeptide operably linked to a polynucleotide sequence encoding a pest control polypeptide, thereby producing the pest resistant plant.