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Isolation and characterization of a cDNA encoding for a lysozyme from the gut of the reduviid bug Triatoma infestans
Author(s) -
Kollien Astrid H.,
Fechner Sabine,
Waniek Peter J.,
Schaub Günter A.
Publication year - 2003
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.10090
Subject(s) - biology , complementary dna , lysozyme , microbiology and biotechnology , triatoma infestans , cdna library , midgut , rapid amplification of cdna ends , peptide sequence , gene , biochemistry , molecular cloning , botany , larva , parasite hosting , trypanosoma cruzi , world wide web , computer science
We have isolated and characterised a Triatoma infestans cDNA encoding a lysozyme. A 174‐bp fragment was amplified by PCR using degenerate oligodeoxyribonucleotide primers derived from the known amino acid sequences of lysozyme from other insects. This PCR fragment was used to screen a cDNA gut library of T. infestans . A clone containing the 3′‐end of the lysozyme cDNA (219 bp) was isolated and sequenced. RACE was used to amplify the 5′‐end of the lysozyme cDNA. After sequencing the complete lysozyme cDNA, the deduced 417 amino acid sequence showed high identity (40–50%) with other chicken‐type lysozymes. The amino acid residues responsible for the catalytic activity and the binding of the substrate were essentially conserved. The expression pattern of the lysozyme gene in bugs at different molting and feeding states showed that this gene was upregulated in the digestive tract directly after the molt and after feeding. Additionally, this lysozyme gene was expressed differently in the different regions of the digestive tract, strongly in the cardia and stomach, the anterior regions of the midgut, and only traces of lysozyme mRNA could be detected in the small intestine, the posterior region of the midgut. Arch. Insect Biochem. Physiol. 53:134–145, 2003. © 2003 Wiley‐Liss, Inc.