Novel Bacterial Proteins with Pesticidal Activity

Abstract

Provided are pesticidal, particularly insecticidal, proteins which resemble S-layer proteins, as well as variants or mutants thereof, and DNAs encoding them. Further provided are methods and means for using said DNA or protein for controlling pests, particularly plant insect pests.

1 . An isolated pesticidal protein characterized by:
 a) being specifically pesticidal to some pests and not to others, and   b) having at least 50% sequence identity with a bacterial S-layer protein.   
     
     
         2 . The protein of  claim 1  which is characterized by:
 a) being specifically insecticidal to some insects and not to others, 
 b) having at least 70% sequence identity to a bacterial S-layer protein, 
 c) a molecular weight of about 50 to about 120 kDa. 
 
     
     
         3 . The protein of  claim 1  which has at least 50% sequence identity to the protein of SEQ ID NO:2 or a toxic fragment thereof, and which has a molecular weight of about 50 to about 120 kDa. 
     
     
         4 . The protein of  claim 2 , which has at least 50% sequence identity to the protein of SEQ ID NO:2 or a toxic fragment thereof, and which has a molecular weight of about 50 to about 120 kDa. 
     
     
         5 . The protein of  claim 4 , comprising the amino acid sequence of SEQ ID NO: 2 from an amino acid position between amino acid position 1 and amino acid position 31 to amino acid position 863. 
     
     
         6 . The protein of  claim 4 , comprising the amino acid sequence of SEQ ID NO: 2 from an amino acid position between amino acid position 1 and amino acid position 531 to amino acid position 863. 
     
     
         7 . The protein of  claim 4  comprising the amino acid sequence of SEQ ID NO: 2. 
     
     
         8 . An isolated DNA encoding the protein of any one of  claims 1  to  7 . 
     
     
         9 . A chimeric gene comprising:
 a) a coding sequence comprising the DNA of  claim 8 , and   b) a promoter which allows expression in plant cells.   
     
     
         10 . The chimeric gene of  claim 9 , wherein said coding sequence is a synthetic DNA sequence that has been optimized for expression in a host plant. 
     
     
         11 . The chimeric gene of  claim 10 , wherein said host plant is selected from the group consisting of: corn, cotton, soybean, rice, oilseed rape, cauliflower, and cabbage. 
     
     
         12 . A plant transformation vector comprising the chimeric gene of any one of  claims 9  to  11 . 
     
     
         13 . A transgenic plant, seed or plant cell comprising the chimeric gene of any one of  claims 9  to  11 . 
     
     
         14 . The plant, seed or cell of  claim 13 , wherein said plant is selected from the group of: corn, cotton, soybean, rice, oilseed rape, cauliflower, and cabbage. 
     
     
         15 . A method of protecting plants against damage caused by plant pests, such as insect pests, feeding on the plant species to which said plants belong, comprising the step of expressing the chimeric gene of any one of  claims 9  to  11  in cells of said plants. 
     
     
         16 . A method of protecting a plant against damage caused by plants pests, such as insect pests, feeding on the plant species to which said plant belongs, comprising the step of transforming a plant cell with the chimeric gene of any one of  claims 9  to  11 , regenerating said cell into a plant, and obtaining progeny and propagating material of said plant, such as seeds, comprising said chimeric gene. 
     
     
         17 . A method for killing pests, comprising contacting said pests with the protein of any one of  claims 1  to  7 . 
     
     
         18 . The method of  claim 17 , wherein said pest is an insect pest. 
     
     
         19 . Use of the DNA of  claim 8  to control or kill pests such as insect pests. 
     
     
         20 . Use of the chimeric gene of any one of  claims 9  to  11  to control or kill pests such as insect pests. 
     
     
         21 . Use of the protein of any one of  claims 1  to  7  to control or kill pests, such as insect pests. 
     
     
         22 . A method to protect a field of plants from pests such as insects, comprising: applying the protein of any one of  claims 1  to  7  to a field of plants, either in the form of a pesticidal composition comprising the protein of any one of  claims 1  to  7 , or in the form of a recombinant organism expressing said protein. 
     
     
         23 . The method of  claim 21  wherein said recombinant organism is a transgenic plant expressing the protein of any one of  claims 1  to  7 .