Abstract: This study aims to clone, sequence and bioinformatics analysis of the entomopathogenic fungus Isaria fumosorosea chalcone isomerase gene. The results of sequence analysis shows that the coding region of IfCHI gene are 1 152 base pairs, encoding 383 amino acids, and the theoretical isoelectric point（pI） is 9.72. It is an unstable water-soluble protein, and the secondary structure is mixed. Its protein solvent accessibility are mainly divided into three categories. The tertiary structure consists of alpha helix, beta sheet, beta turn and random coil and there are significant differences between homologous sequence analysis and multiple sequence alignment analysis. The successful cloning and analysis of the insect pathogen of IfCHI coding region will provide a foundation for further study of the genetic characteristics and expression mechanism of the pathogen CHI gene, and search for more chalcone compounds.