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'); Reports of Biochemistry and Molecular Biology rbmb.net Basic Sciences http://rbmb.net 1 admin 2322-3480 2322-3480 10.61882/rbmb en jalali 1391 7 1 gregorian 2012 10 1 1 1 online 1 fulltext
en Cloning and Expression of Cyclophilin from Platanus orientalis Pollens in Escherichia coli بیوشیمی Biochemistry <p style="text-align: justify;"><em><strong>Background:</strong></em> Allergy is a clinical disorder affecting the human population with wide geographical distribution. <em>Platanus orientalis (P. orientalis)</em> trees are planted in many countries and their pollen causes allergic reactions.</p> <p style="text-align: justify;">Cyclophilin has recently been identified as one of the most important allergens of<em> P. orientalis</em> pollen. We aimed to clone and purify this allergen in <em>Escherichia coli </em>for further studies and therapeutic and diagnostic purposes for allergy to<em> P. orientalis.</em></p> <p style="text-align: justify;"><em><strong>Methods:</strong></em> RNA was extracted from <em>P. orientalis.</em> A full-length fragment encoding <em>cyclophilin</em> was prepared by polymerase chain reaction amplification of the first-strand cDNA synthesized from <em>P. orientalis</em> RNA. The cDNA was inserted into the pET32b (+) vector, and the construct transformed into <em>E. coli</em> Top10 and BL21 cells. The expressed protein was purified by the CuSO4 method.</p> <p style="text-align: justify;"><em><strong>Results:</strong></em> The cDNA for the cyclophilin of <em>P. orientalis</em> pollen was cloned, and a specific reactivity of recombinant cyclophin was confirmed by immunoblotting using sera from patients allergic to <em>P. orientalis</em> pollen.</p> <p style="text-align: justify;"><em><strong>Conclusion:</strong></em> The recombinant cyclophilin has a potential for immunologic assays for evaluation of allergy to <em>P. orientalis</em> pollen.</p> Allergy, Cloning, Cyclophilin, Escherichia coli, Platanus orientalis, Pollen, Recombinant allergen 25 29 http://rbmb.net/browse.php?a_code=A-10-1-5&slc_lang=en&sid=1 Mojtaba Sankian 10031947532846005901 10031947532846005901 No Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran Fatemeh Vahedi 10031947532846005902 10031947532846005902 No Razi Vaccine & Serum Research Institute, Mashhad, Iran Nazanin Pazouki 10031947532846005903 10031947532846005903 No Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran Malihe Moghadam 10031947532846005904 10031947532846005904 No Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran Farahzad Jabbari Azad 10031947532846005905 10031947532846005905 No Allergy Research Center, Mashhad University of Medical Sciences, Mashhad, Iran Abdol-Reza Varasteh varasteha@mums.ac.ir 10031947532846005906 10031947532846005906 Yes Allergy Research Center, Mashhad University of Medical Sciences, Mashhad, Iran