In this study, the trypsin gene (bgtryp-1) from the German cockroach, Blattella germanica, was cloned via the immunoscreening of patients with allergies to cockroaches. Nucleotide sequence analysis predicted an 863 bp open reading frame which encodes for 257 amino acids. The deduced amino acid sequence exhibited 42-57% homology with the serine protease from dust mites, and consisted of a conserved catalytic domain (GDSGGPLV). bgtryp-1 was determined by both Northern and Southern analysis to be a 0.9 kb, single-copy gene. SDS-PAGE and Western blotting analyses of the recombinant protein (Bgtryp-1) over-expressed in Escherichia coli revealed that the molecular mass of the expressed protein was 35 kDa, and the expressed protein was capable of reacting with the sera of cockroach allergy patients. We also discussed the possibility that trypsin excreted by the digestive system of the German cockroach not only functions as an allergen, but also may perform a vital role in the activation of PAR-2.
Allergens/analysis/chemistry/*genetics ; Amino Acid Sequence ; Base Sequence ; Blattellidae/*genetics ; Blotting, Western ; Catalytic Domain/genetics ; DNA, Complementary/genetics ; Escherichia coli/metabolism ; Female ; *Genes, Insect ; Humans ; Male ; Molecular Sequence Data ; Molecular Weight ; Receptor, PAR-2/metabolism ; Recombinant Proteins/analysis/biosynthesis/chemistry ; Research Support, Non-U.S. Gov't ; Sequence Alignment ; Sequence Homology, Amino Acid ; Trypsin/analysis/chemistry/*genetics