Insecticidal proteins

A method of making an engineered hybrid insecticidal protein (eHIP), comprising:

• a. obtaining a first nucleic acid that encodes a Bacillus thuringiensis (Bt) Cry3A protein;
• b. obtaining a second nucleic acid that encodes a Bt Cry1Aa or Cry1Ab protein;
• c. fusing in an 5′ to 3′ direction a 5′ portion of the first nucleic acid that encodes an N-terminal region of the Cry3A protein to a 3′ portion of the second nucleic acid that encodes a C-terminal region of the Cry1Aa or Cry1Ab protein, wherein at least one crossover position between the first and the second nucleic acids is located in the nucleotides that encode conserved block 2, conserved block 3 or variable region 4 to make a hybrid nucleic acid that encodes an eHIP that has activity against western corn rootworm, wherein the engineered insecticidal protein has at least 80% identity to SEQ ID NO:64;
• and optionally fusing to the 5′ end of the hybrid nucleic acid a nucleotide sequence that encodes a peptidyl fragment resulting in a 5′ extension or fusing to the 3′ end of the hybrid nucleic acid a nucleotide sequence that encodes a protoxin tail region of a Bt Cry protein resulting in a 3′ extension, or both;
• d. inserting the hybrid nucleic acid with or without one or both of the 5′ or 3′ extensions into an expression cassette;
• and
• e. transforming the expression cassette into a host cell, resulting in the host cell producing an engineered hybrid insecticidal protein.