Molecular cloning of a small heat shock protein (sHSPII) from the cattle tick Rhipicephalus (Boophilus) annulatus salivary gland

作者：Shahein Yasser E^*; Abd El Rahim Mohamed T; Hussein Nahla A; Hamed Ragaa R; El Hakim Amr E; Barakat Maged M

来源：International Journal of Biological Macromolecules, 2010, 47(5): 614-622.

DOI：10.1016/j.ijbiomac.2010.08.006

• Rhipicephalus annulatus
• sHSP
• Protective immunogens
• Thermal stability
• Salivary glands

Immunoscreening of a cDNA expression library of the Rhipicephalus (Boophilus) annulatus tick with purified rabbit anti-R annulatus salivary glands antigens polyclonal antibodies led to the identification of a 661 bp sequence The sequence includes an open reading frame of 543 bp encoding a protein of 180 amino acids with calculated molecular weight of 20 51 kDa isoelectric point of 9 071 and with no signal sequence Comparison of the deduced amino acids with protein data bank showed that the identified polypeptide belongs to the alpha crystallin small heat shock proteins superfamily and shows sequence similarity of 62% and 55% to Ixodes scapulans fed tick salivary gland protein and Ornithodoros parken alpha-crystallin protein respectively Accordingly this protein was called Ra-sHSPII The Ra-sHSPII protein was expressed in E coli under T7 promotor of the pET-30b vector purified under denaturation conditions and the immunogenicity and cross-reactivity of the recombinant Ra-sHSPII were evaluated Direct ELISA showed that the Ra-sHSPII is a strong immunogen In immunoblotting assay the anti-rRa-sHSPII antisera reacted specifically with purified rRa-sHSPII with several proteins in R. annulatus whole tick larval and gut protein extracts in addition to Hyalomma dromedaru and Ornithodoros moubata whole tick protein extracts as examples of hard and soft tick species respectively The rRa-sHSPII protein confers thermal protection to other proteins in vitro as found in other sHSPs E coli cell extracts containing the prote

• 出版日期2010-12-1