Insect pests are one of the major biotic factors limiting the yield of agricultural food crops. In routine agricultural practice, chemical pesticides are used to control insect pests; however, repeated spraying leads to numerous environmental and health concerns. The search for eco-friendly and sustainable alternatives to chemical pesticides has led to the exploitation of biological control agents. Among these, chitinase plays a target-specific insecticidal activity by degrading the chitin in the insect exoskeleton and gut regions, making it a desirable alternative to chemical pesticides. Therefore, the objective of the study was to isolate and clone the Pseudomonas fluorescens chitinase gene for its use as a specific biopesticide. Full-length chitinase gene coming under family 18 Group D glycosyl hydrolases was isolated from P. fluorescens genomic DNA using polymerase chain reaction amplification and cloned into an expression vector pET32C+ and transformed into Escherichia coli. The sequencing results showed that the chi gene contained an 1800 bp long open reading frame encoding 353 amino acids. The deduced amino acid sequence showed that the protein consisted of the chitin-binding domain, a catalytic domain, and a fibronectin Type III domain also an amino terminus signal peptide. This study allows the identification of new, target-specific bacterial metabolite as a biopesticide for safe, environment-friendly pest management strategies.
Key words: Agricultural practice, biopesticide, chitin, cloning, domains, expression vector, Pest management.
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