A new insertion variant belonging to the family IS231, designated IS231I, was isolated from a mosquito larvicidal strain of the Bacillus thuringiensis serovar sotto (H4ab). IS231I was 1653 bp long and delimited by two 20 bp inverted repeats with one mismatch, flanked by two perfect 11 bp direct repeats. The element contained a single open reading frame (ORF) encoding 478 amino acids and five conserved domains: N1, N2, N3, C1, and C2. The 5' noncoding region upstream of the ORF, presumed to form a stable stem-and-loop structure, was highly conserved in IS231I. The secondary structure conformation had a deduced free energy (Delta G = 25 degrees C) of -17.2 kcal/mol. Comparison of the IS231I amino acid sequence with those of the 10 existing IS variants revealed that the new variant shares 89% identity with IS231A and IS231B, 65-66% with IS231M and IS231N, and 38% with IS231W.