The invention provides a novel Bt protein Cry4Cb2 and a coding gene thereof, wherein the protein has an amino acid sequence shown as SEQ ID No.2 or an amino acid sequence which is shown as the SEQ ID No.2, is subjected to substitution, deletion and/or increasing one or more amino acids, and has protein with the same activity. The protein can be used for preparing Bt pesticides, and the gene can convert crops such as cotton, corn, rice, vegetables and the like and make the crops have corresponding anti-insect activity, thereby reducing the using amount of pesticides, reducing environmental pollution, and having important economic value and application prospect.