www.elsevier.com/locate/ibmb Trypsinogen-like cDNAs and quantitative analysis of mRNA levels from the Indianmeal moth, Plodia interpunctella

Two cDNA fragments encoding full-length trypsinogen-like proteins were cloned from larvae of two strains (RC688s and HD198r) of the Indianmeal moth, Plodia interpunctella (Hübner), which differed in their sensitivity to Bacillus thuringiensis protoxins. One cDNA fragment contained 874 nucleotides, including a 780-nucleotide open reading frame that encoded a trypsinogen-like protein (PiT2b). Another cDNA fragment amplified from both P. interpunctella strains contained 864 nucleotides including a 780 bp open reading frame encoding a second trypsinogen-like protein (PiT2c). The cDNA sequence of PiT2b shared 89 % sequence identity with PiT2a, a trypsinogen-like protein cloned previously from this species. The cDNA sequences of PiT2a and PiT2c shared 83% identity. The cDNA sequence identity between PiT2b and PiT2c was 80%. The cDNA for PiT2b from strain RC688s was different at six nucleotide positions from that of PiT2b from strain HD198r. Five nucleotide replacements occurred in the open reading frame leading to amino acid changes at all five positions. There were five nucleotide differences in the cDNAs for PiT2c trypsinogen-like proteins from the two strains. Two nucleotide substitutions in the open reading frame resulted in replacements of two amino acid residues in the deduced protein sequences. Amino acid sequences for PiT2a and PiT2b shared 84 % identity, but only 50% identity was observed between PiT2c and the other two trypsinogen-like proteins. The deduced amino acid sequences for PiT2b and PiT2c included both signal and zymogen activation peptides and amino acid sequence motifs which are conserved in seven homologous trypsinogen-like proteins from other insects. Typical features of the putative trypsinogen-like proteins from P. inter