Document Details

Document Type : Article In Journal 
Document Title :
Molecular cloning of a small heat shock protein (sHSPII) from the cattle tick
الإستنساخ الجزيئي لبروتين الصدمة الحرارية الصغير من الغدة اللعابية لقراد البقر من نوع ريبيسيفيلس (بوؤفيلس) أنيولاتس
 
Subject : Biochemistry 
Document Language : English 
Abstract : 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 scapularis fed tick salivary gland protein and Ornithodoros parkeri alphacrystallin 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 dromedarii 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 protein were protected from heat-denatured precipitation when heated up to 100 ◦C, whereas extracts from cells not expressing the protein were heat-sensitive at 60 ◦C. 
ISSN : 8130-0141 
Journal Name : International Journal of Biological Macromolecules 
Volume : 47 
Issue Number : 5 
Publishing Year : 1431 AH
2010 AD 
Article Type : Article 
Added Date : Saturday, January 15, 2011 

Researchers

Researcher Name (Arabic)Researcher Name (English)Researcher TypeDr GradeEmail
عمرو السيد الحكيمEl Hakim, Amr El SayedResearcherDoctorateaselhakim@hotmail.com
ماجد محمد بركاتBarakat, Maged MohamedResearcherDoctorate 
نهلة عبد المعطي حسينHussein, Nahla Abdel MoetyResearcherMaster 
ياسر عزت شاهينShahein, Yasser EzzatResearcherDoctorate 
محمد طه عبد الرحيمAbdel Rahim, Mohamed TahaResearcherDoctorate 
رجاء رضا حامدHamed, Ragaa RedaResearcherDoctorate 

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