title: Identification of the heat shock protein 70 (HLHsp70) in Haemaphysalis longicornis.
abstract: A Haemaphysalis longicornis heat shock protein 70 (HLHsp70) was identified from a cDNA library synthesized from tick eggs. The HLHsp70 cDNA is 2311 bp in length and encodes 661 amino acid residues with the predicted molecular weight of 72.5 kDa and an isoelectronic point (pI) of 5.2. It also contains the highly conserved functional motifs of the Hsp70 family and a specific endoplasmic reticulum (ER) retention signal "KDEL" that is common among ER-localized proteins. The HLHsp70 exhibits 90% amino acid identity to the putative Hsp70 of Ixodes scapularis, and 85% to Gallus gallus 78 kDa glucose-regulated protein precursor. Real time RT-PCR analysis showed that the expression levels of the Hsp70 in ovaries and salivary glands were significantly higher than in other tested tissues in partially fed females. Although the expression level of the HLHsp70 was constantly low in unfed ticks, it was significantly induced by blood-feeding. Further, the expression was positively correlated to the temperature (4-37°C, tested). Western blot analysis showed that the rabbit antiserum against the recombinant HLHsp70 protein (rHLHSP70) recognized bands of approximately 100, 72, and 28 kDa from egg lysates, as well as a 72kDa fragment in protein extracts from partially fed larvae. Immunization of rabbits with the rHLHSP70 did not result in a statistically significant reduction of female tick engorgement and oviposition. These results suggest that although HLHSP70 plays a role in the physiological activities of ticks, as a constitutive protein it was not suitable for selection as a candidate vaccine antigen against ticks.
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 1 Xujianping, Yanchangbu, Lanzhou, Gansu Province 730046, People's Republic of China.
authors.forename: Zhancheng
authors.initials: Z
authors.lastname: Tian
authors.forename: Guangyuan
authors.initials: G
authors.lastname: Liu
authors.forename: Liyan
authors.initials: L
authors.lastname: Zhang
authors.forename: Hong
authors.initials: H
authors.lastname: Yin
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.forename: Junren
authors.initials: J
authors.lastname: Xie
authors.forename: Ping
authors.initials: P
authors.lastname: Zhang
authors.forename: Jin
authors.initials: J
authors.lastname: Luo
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D019295:Computational Biology; D004064:Digestive System / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation; D015723:Gene Library; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D007814:Larva / Q000378:metabolism; D008969:Molecular Sequence Data; D009758:Nymph / Q000378:metabolism; D010053:Ovary / Q000378:metabolism; D010063:Ovum / Q000378:metabolism; D010802:Phylogeny; D011817:Rabbits; D012469:Salivary Glands / Q000378:metabolism; D013987:Ticks / Q000235:genetics / Q000378:metabolism*
chemical_list: D018840:HSP70 Heat-Shock Proteins