Abstract
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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Acknowledgments
This work was supported by a grant from the Bio-oriented Technology Research Advancement Institution (BRAIN), Grants-in-Aid for Scientific Research (A) from the Japan Society for the Promotion of Science, and a grant from the 21st Century COE program (A-1), Ministry of Education, Sports, Science, and Technology of Japan.
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Zhou, J., Liao, M., Gong, H. et al. Characterization of Hlcyst-3 as a member of cystatins from the tick Haemaphysalis longicornis . Exp Appl Acarol 51, 327–333 (2010). https://doi.org/10.1007/s10493-010-9336-1
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DOI: https://doi.org/10.1007/s10493-010-9336-1