Cloning and expression of cry2Aa from native Bacillus thuringiensis strain SY49-1 and its insecticidal activity against Culex pipiens (Diptera: Culicidae)

Abstract Bacillus thuringiensis (Berliner) ( Bt ) is well known for having toxicity against pest insects because of their ability to form endospores and broad-range activity of their parasporal inclusions. In this study, a new member of cry2A gene from previously characterized native B. thuringiensis SY49-1 strain was cloned, expressed and used for its activity against Culex pipiens (Diptera: Culicidae) larvae. The sequence analysis of the cloned cry2A gene revealed that it encodes a polypeptide of 633 aa residues with 99% identity to Cry2Aa protein with expected molecular weight of 70.7 kDa. Bacillus thuringiensis delta-endotoxin nomenclature committee designed our sequence as Cry2Aa18 being a new member of Bt toxins. Bioassays against last instar larvae of C. pipiens indicated that Cry2Aa18 has considerable toxicity with LC 50 of 630 μg ml −1 . In order to prevent the spread of infectious diseases mediated by C . pipiens , this newly characterized cry2Aa18 gene could constitute as an important biological control tool for controlling mosquito larvae living in freshwater systems and can be used as a good alternative for minimizing the use of chemicals.