Novel bacillus thuringiensis insecticidal proteins

Abstract

The invention pertains to novel insecticidal compounds derived from Bacillus thuringiensis strains. New proteins designated Cry2Ae, Cry2Af, and Cry2Ag, and variants thereof are provided, as well as DNA sequences encoding these proteins or their variants. Further provided are recombinant hosts expressing such proteins, particularly plant cells and plants.

1 - 56 . (canceled) 
     
     
         57 . A method for controlling  Helicoverpa armigera, Anticarsia gemmatalis  or  Sesamia inferens  comprising applying an insecticidal protein to an area or plant to be protected, wherein said area or plant is a habitat of  Helicoverpa armigera, Anticarsia gemmatalis  or  Sesamia  inferens, and wherein said insecticidal protein is
 i) an insecticidal protein comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO. 2,   ii) an insecticidal protein comprising the amino acid sequence from the amino acid at position 1 to the amino acid at position 625 in SEQ ID No. 2,   iii) an insecticidal protein comprising the amino acid sequence from the amino acid at position 50 to the amino acid at position 625 in SEQ ID No. 2,   iv) an insecticidal protein comprising the amino acid sequence of SEQ ID No. 2,   v) an insecticidal protein having 5-10 amino acids added, replaced or deleted in the sequence of SEQ ID NO: 2 without changing the insecticidal activity of the protein,   vi) an insecticidal protein having less than 5 amino acids added, replaced or deleted in the sequence of SEQ ID NO: 2 without changing the insecticidal activity of the protein,   vii) an insecticidal protein comprising the amino acid sequence of SEQ ID No. 8, or   viii) an insecticidal protein encoded by the DNA of SEQ ID No 7 or 9.   
     
     
         58 . The method of  claim 57 , wherein said applying comprises spraying an insecticidal amount of said insecticidal protein. 
     
     
         59 . The method of  claim 57 , wherein said applying comprises expressing said insecticidal protein in said plant. 
     
     
         60 . The method of  claim 57 , wherein said plant is soybean, rice, corn or cotton. 
     
     
         61 . The method of  claim 59 , wherein said plant is soybean, rice, corn or cotton. 
     
     
         62 . The method of  claim 57 , wherein said plant expresses another insecticidal protein, wherein said another insecticidal protein is a Cry1F protein or a hybrid derived from a Cry1F protein, a Cry1A protein or a toxic fragment thereof, a Cry1Ac protein or a hybrid derived from a Cry1Ac protein, or a VIP3Aa protein or a toxic fragment thereof. 
     
     
         63 . The method of  claim 59 , wherein said plant expresses another insecticidal protein, wherein said another insecticidal protein is a Cry1F protein or a hybrid derived from a Cry1F protein, a Cry1A protein or a toxic fragment thereof, a Cry1Ac protein or a hybrid derived from a Cry1Ac protein, or a VIP3Aa protein or a toxic fragment thereof. 
     
     
         64 . The method of  claim 57 , wherein said plant is transformed with a DNA encoding a protein conferring resistance to a herbicide based on glufosinate or glyphosate. 
     
     
         65 . The method of  claim 59 , wherein said plant is transformed with a DNA encoding a protein conferring resistance to a herbicide based on glufosinate or glyphosate. 
     
     
         66 . The method of  claim 59 , wherein said insecticidal protein has at least 96% sequence identity to the amino acid sequence of SEQ ID NO. 2. 
     
     
         67 . The method of  claim 59 , wherein said insecticidal protein has at least 97% sequence identity to the amino acid sequence of SEQ ID NO. 2. 
     
     
         68 . The method of  claim 59 , wherein said insecticidal protein has at least 98% sequence identity to the amino acid sequence of SEQ ID NO. 2. 
     
     
         69 . The method of  claim 59 , wherein said insecticidal protein has at least 99% sequence identity to the amino acid sequence of SEQ ID NO. 2. 
     
     
         70 . The method of  claim 57 , wherein said insecticidal protein comprises the amino acid sequence from the amino acid at position 1 to the amino acid at position 625 in SEQ ID No. 2, or comprises the amino acid sequence from the amino acid at position 50 to the amino acid at position 625 in SEQ ID No. 2, or comprises the amino acid sequence of SEQ ID No. 2. 
     
     
         71 . The method of  claim 59 , wherein said insecticidal protein comprises the amino acid sequence from the amino acid at position 1 to the amino acid at position 625 in SEQ ID No. 2, or comprises the amino acid sequence from the amino acid at position 50 to the amino acid at position 625 in SEQ ID No. 2, or comprises the amino acid sequence of SEQ ID No. 2. 
     
     
         72 . The method of  claim 57 , wherein said insecticidal protein is an insecticidal protein having 5-10 amino acids added, replaced or deleted in the sequence of SEQ ID No. 2 without changing the insecticidal activity of the protein. 
     
     
         73 . The method of  claim 59 , wherein said insecticidal protein is an insecticidal protein having less than 5 amino acids added, replaced or deleted in the sequence of SEQ ID No. 2 without changing the insecticidal activity of the protein. 
     
     
         74 . The method of  claim 61 , wherein said insecticidal protein comprises the amino acid sequence from the amino acid at position 50 to the amino acid at position 625 in SEQ ID No. 2, or wherein said insecticidal protein is an insecticidal protein having less than 5 amino acids or is an insecticidal protein having 5-10 amino acids added, replaced or deleted in the sequence of SEQ ID No. 2 without changing the insecticidal activity of the protein. 
     
     
         75 . An isolated DNA sequence encoding a truncated insecticidal Cry2Ae protein consisting of the amino acid sequence of the protein of SEQ ID NO: 2 from amino acid position 1 to an amino acid position between amino acid position 625 and amino acid position 632. 
     
     
         76 . A method for controlling insects comprising expressing in transformed plant cells an insecticidally-effective amount of said truncated insecticidal Cry2Ae protein encoded by the DNA of  claim 75 , to control  Heliothis virescens, Helicoverpa zea, Helicoverpa armigera, Anticarsia gemmatalis  and  Ostrinia nubilalis, Chilo suppressalis, Chilo partellus, Scirpophaga incertulas, Sesamia inferens, Cnaphalocrocis medinalis, Marasmia patnalis, Marasmia exigua, Marasmia ruralis , or  Scirpophaga innotata.