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

OCK Mee Sun (Department of Parasitology, Kosin Medical College)
KIM Bong Jin (Department of Parasitology, Kosin Medical College)
KIM Sun Mi (Department of Parasitology, Kosin Medical College)
BYUN Kang Hyun (Department of Parasitology, Kosin Medical College)
Publication Information
Parasites, Hosts and Diseases / v.43, no.3, 2005 , pp. 101-110 More about this Journal
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 (GOSGGPLV). 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 cock-roach 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.
Keywords
Blattella germanica; trypsin; serine protease; allergen; PAR-2;
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