Abstract：This study was based on Bacillus thuringiensis(Bt) strain FH21 with high toxic of Plutella xylostella larvae. Using an improved polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) method, cry1Bb3, cry1Da4, cry1Ja3 and cry1Id3 genes were cloned from strain FH21 with the GenBank No. KJ619659, KJ619660, JQ228425 and KJ619661, respectively. The 4 genes were inserted into expression vector pEB, and transformed into Escherichia coli strain Rosetta (DE3), which were induced by isopropyl β-D-1-thiogalactopyranoside (IPTG) at low temperature. The total protein of FH21 and the 4 proteins were used for bioassay on P. xylostella, Helicoverpa armigera and Ostrinia furnacalis. The total protein of FH21 had high insecticidal activity against 3 insects with lethal concentration 50 (LC50) of 0.801, 0.502 and 2.858 mg/L, respectively. Cry1Bb3, Cry1Ja3 and Cry1Id3 had high insecticidal activity against P. xylostella and O. furnacalis with LC50 of 1.860, 1.697, 1.538, 1.800, 2.270 and 0.840 mg/L, respectively. This study showed that Bt FH21strain against agricultural pests with a high insecticidal activity, insecticidal genes were assayed and insecticidal activity were determined simultaneously. The cry1 genes provide more choices for important agricultural pest.
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