Korean J Parasitol.  2005 Sep;43(3):101-110. 10.3347/kjp.2005.43.3.101.

Cloning and expression of trypsin-encoding cDNA from Blattella germanica and its possibility as an allergen

• Ock MS
• Kim BJ
• Kim SM
• Byun KH

• KMID: 1720500
• DOI: http://doi.org/10.3347/kjp.2005.43.3.101

Abstract

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.

Keyword

Blattella germanica; trypsin; serine protease; allergen; PAR-2

MeSH Terms

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