title: Molecular cloning of Ra-sHSPI, a novel member of the HSP20 family from Rhipicephalus annulatus salivary glands.
abstract: Infestation of cattle by ticks of Rhipicephalus spp. results in severe veterinary and economical losses. Identification of novel proteins from tick salivary glands will enhance our understanding of several aspects of tick physiology and will aid in the development of anti-tick vaccines. Small heat shock proteins (HSPs) have important roles in infection and immunity, especially between invertebrate vectors and mammalian hosts while initially performing their molecular chaperone activity. Here, we report the identification of a small HSP gene from the salivary glands of Rhipicephalus annulatus ticks through immunoscreening of the corresponding cDNA expression library. The identified cDNA contained a 742bp sequence with 543bp open reading frame. It was subsequently cloned, expressed and successfully purified under both native and denaturing conditions. Sequence analysis and functional investigations showed that the protein belongs to the HSP20 family, hence the annotated name Ra-sHSPI. Indeed, recombinant Ra-sHSPI showed two typical in vitro activities of holdase chaperones, including thermal protection of bacterial cellular extracts and the recombinant HindIII at elevated temperatures. Moreover, the recombinant Ra-sHSPI showed strong immunogenic effect in animal model. These results pave the way toward further investigation of Ra-sHSPI role in ticks feeding and its potential use as protective antigen.
authors.affiliation: Molecular Biology Department, Genetic Engineering and Biotechnology Division, National Research Centre, Dokki, Cairo, Egypt.
authors.forename: Nahla A
authors.initials: NA
authors.lastname: Hussein
authors.affiliation: Molecular Biology Department, Genetic Engineering and Biotechnology Division, National Research Centre, Dokki, Cairo, Egypt. Electronic address: yassershahein_nrc@yahoo.com.
authors.forename: Yasser E
authors.initials: YE
authors.lastname: Shahein
authors.affiliation: Molecular Biology Department, Genetic Engineering and Biotechnology Division, National Research Centre, Dokki, Cairo, Egypt.
authors.forename: Amr E
authors.initials: AE
authors.lastname: El-Hakim
authors.affiliation: Radiation Biology Department, National Centre for Radiation Research and Technology (NCRRT), Nasr City, Cairo, Egypt.
authors.forename: Amira M
authors.initials: AM
authors.lastname: Abouelella
authors.affiliation: Molecular Biology Department, Genetic Engineering and Biotechnology Division, National Research Centre, Dokki, Cairo, Egypt.
authors.forename: Rasha A
authors.initials: RA
authors.lastname: Guneidy
authors.affiliation: Molecular Biology Department, Genetic Engineering and Biotechnology Division, National Research Centre, Dokki, Cairo, Egypt.
authors.forename: Ragaa R
authors.initials: RR
authors.lastname: Hamed
mesh_terms: D000818:Animals; D002417:Cattle / Q000235:genetics / Q000469:parasitology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D050886:HSP20 Heat-Shock Proteins / Q000235:genetics*; D048494:Rhipicephalus / Q000235:genetics*; D012469:Salivary Glands; D017422:Sequence Analysis, DNA
chemical_list: D018076:DNA, Complementary; D050886:HSP20 Heat-Shock Proteins
keywords: Cattle ticks; Rhipicephalus annulatus; Salivary glands; Thermal stability; sHSP