Journal
EXPERIMENTAL AND APPLIED ACAROLOGY
Volume 51, Issue 4, Pages 327-333Publisher
SPRINGER
DOI: 10.1007/s10493-010-9336-1
Keywords
Haemaphysalis longicornis; Cystatin; Hlcyst-3
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Funding
- Bio-oriented Technology Research Advancement Institution (BRAIN)
- Japan Society for the Promotion of Science
- Ministry of Education, Sports, Science, and Technology of Japan [A-1]
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The gene encoding cystatin from the tick Haemaphysalis longicornis has been reported previously. In the study reported here, we characterized a member of cystatins and designated it as Hlcyst-3 (H. longicornis cystatin-3). Its full-length cDNA is 602 bp, and it encodes a putative 129 amino acid protein with an obvious signal peptide. Sequence analysis revealed that it has significant homology with the known secreted cystatin. The recombinant protein was expressed in a GST-fused soluble form in Escherichia coli and was purified by affinity chromatography. The inhibitory activity of the recombinant protein against papain and cathepsin L was identified by fluorogenic substrate analysis. Real-time PCR revealed that Hlcyst-3 was mostly expressed in the tick midgut.
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